Bergamo® II Series Multiphoton Microscopes

Overview
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Configurations
Retrofits
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Sam Rubin
Imaging Systems General Manager

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Imaging Systems Demo Rooms

To ensure our customers choose the best imaging system for their needs, Thorlabs' operates showrooms at a number of locations worldwide.

Click on the locations to the right for contact information and available demo systems.

Two Scientists Using a Bergamo II Microscope

Try Our Microscopes in Person

The Bergamo^® II Series is Thorlabs' platform for multiphoton microscopy. Following the principle that the microscope should conform to the specimen, rather than the other way around, we created a completely modular imaging platform that adapts to a wide range of experimental requirements and can be easily upgraded as experimental needs evolve.

Options at a Glance

Laser Scanning

8 kHz and 12 kHz Resonant-Galvo-Galvo and Galvo-Resonant Scanners for High-Speed Imaging
Galvo-Galvo Scanners for User-Defined ROI Shapes and Photostimulation Patterns
Spatial Light Modulator (SLM) for Simultaneous Multi-Point Targeting
Super Broadband Scan Optics Optimized for:
- Photoactivation / Uncaging
- Two-Photon Imaging
- Three-Photon Imaging
Co-Registered Confocal Imaging

Microscope Body

Rotating Bodies (See Video to the Lower Right)
- 5" of Coarse Vertical Motion
- -5° to +95° or -50° to +50° Rotation Around the Sample (-45° to +45° Only with SLM)
- 2" of Fine XY Motion
- 1" of Fine Z Motion
- X, Y, and Z Rotate with Objective
Upright Bodies
- Fine Z or Fine XYZ Motion

Signal Detection

Up to Four Simultaneous Detection Channels
Thermoelectrically Cooled or Large-Angle Non-Cooled GaAsP and Multialkali PMTs
8°, 10°, or 14° Collection Optics in Epi Direction (for Ø20 mm Entrance Pupil)
13° in Transmission Direction
Mechanical Shutters Available for Photostimulation
Option for Two Forward-Direction Detection Channels
Easy-to-Exchange Magnetic Filter Holders

Transmitted Light Imaging

Dodt Gradient Contrast (Widefield and Laser Scanned)
DIC (Widefield and Laser Scanned)
Illumination Modules are Easily Removed for In Vivo Experiments
Visible and NIR LED Illumination
Available for Rotating and Upright Bodies

Volume Imaging Using Bessel Beams

3D Video-Rate for Volumetric Functional Imaging
Enhanced Temporal Resolution Adequate for Studying Internal Systems at Cellular Lateral Resolution In Vivo
Want to Learn More? Press Release

Innovation through Collaboration

Five-Axis Movement of Rotating Bergamo Microscopes

Features

The features of Bergamo^® II listed below reflect our focus on developing cutting-edge capabilities without compromising usability. Many of these features can also be added to existing Thorlabs microscope systems. For more information, please see the Retrofits tab.

Laser Scanning, Widefield Imaging, and Transmitted Light Imaging
Scan Paths	Resonant-Galvo-Galvo	8 kHz Scanner: Image at 2 fps (4096 x 4096 Pixels), 30 fps (512 x 512 Pixels), or 400 fps (512 x 32 Pixels) 12 kHz Scanner: Image at 45 fps (512 x 512 Pixels) or 600 fps (512 x 32 Pixels)
	Galvo-Resonant	8 kHz Scanner: Image at 2 fps (4096 x 4096 Pixels), 30 fps (512 x 512 Pixels), or 400 fps (512 x 32 Pixels) 12 kHz Scanner: Image at 45 fps (512 x 512 Pixels) or 600 fps (512 x 32 Pixels)
	Galvo-Galvo	User-Defined Scan Geometries: Squares, Rectangles, Circles, Ellipses, Lines, and Polylines Capture Weak Signals with Long Dwell Time Integration Consistent Dwell Times Across Field of View 48 fps at 512 x 32 Pixels and 70 fps at 32 x 32 Pixels
	Spatial Light Modulator	Simultaneous Multi-Site Photostimulation using Holographic Beam Control Entirely Controlled Through ThorImage^®LS Software Precise Z-Axis Control of Photoexcited Locations
Widefield Viewing		Compatible with Thorlabs or Third-Party C-Mount-Threaded Scientific Cameras Locate Areas of Interest without Unnecessary Laser Excitation
Epi-Illumination		Single-Filter-Cube or Six-Filter-Turret Epi-Illuminator Modules Visualize Samples with Fluorescence or Reflected Light Variety of Available Sources for Brightfield Illumination Thorlabs' Mounted and Solis™ LEDs Optional Port for Industry-Standard Liquid Light Guides
Dodt Gradient Contrast and DIC Modules		User-Removable Modules Convert Microscope Between In Vivo and Slice Imaging Widefield Imaging or Laser Scanning Dodt Gradient Contrast: View Features in Tissue Slices DIC: View Features in Thinner, Transparent Samples

Optical Performance
PMT Configuration		Choice of Signal Collection Angles to Accommodate Different Sample Depths along Epi Path 8° (For Two PMTs)^a 10° (For up to Four PMTs)^a 14° (For Two PMTs)^a Option for Highly Sensitive Forward Fluorescence Detection Channels along Transmitted Path 13° (For up to Two PMTs)^a GaAsP and Multialkali PMTs Available All PMTs Available with or without Mechanical Shutters for Photoactivation Experiments Angles Quoted for an Objective with a Ø20 mm Entrance Pupil
Minimal Distance Between Objective and First Collecting Lens		Large Collection Angle for Multiphoton Fluorescence Emission Increased Collection Efficiency
Periscopes		Maintain Laser Alignment and Optical Performance over Microscope's Entire Travel Range
Scan Paths Designed In-House		Super Broadband Correction Range of 450 - 1100 nm, 680 - 1600 nm, or 900 - 1900 nm Optimized for Photostimulation, Two-Photon Imaging, or Three-Photon Imaging Specifically Designed to Match the Low-Magnification, High-NA Objectives Popularly Used in Multiphoton Microscopy Take Full Advantage of the Latest Widely Tunable Ti:Sapphire and OPO Systems Fill up to a Ø20 mm Back Aperture Objective Up to F.N. 20

Day-to-Day Usage
Several Software Packages		ThorImage^®LS: Our Internally Developed, Open-Source Solution Full SDK for LabVIEW and C++ Available Upon Request Compatible with ScanImage
Touchscreen Controller		Touchscreen Shows Current Position of All Axes Tap to Save and Retrieve Two Positions in Space
Easy-to-Reach Emission Filters and Dichroic Holders		Filters are Accessed from the Front of the Microscope and Take Less than 5 Minutes to Exchange
Input and Output Triggers		Use Electrical Signals to Synchronize All Your Equipment Input Triggers Can Start a Single Series or an Indefinite Series Output Triggers Can be Sent at the Beginning of a Frame or Line High-Bandwidth Signal Integration with Electrophysiology
Large User-Adjustable Volume Underneath Objective		Accommodates Large Preps and Setups Approach Angles Around Objective are Not Restricted Rotating Bodies have 5" (12.7 cm) of Elevator Travel for Easily Adapting the Microscope for Differently Sized Experimental Setups
≥90° Rotation of the Focal Plane (Rotating Bodies Only)		Image Different Sections of the Brain without Having to Move Your Specimen or Refocus the Objective Rotation Angles: -5° to +95° or -50° to +50° for Microscopes without SLM -45° to +45° for Microscopes with SLM

Beam Conditioning Modules
Pockels Cells		Edge and Fly-Back Blanking to Minimize Sample Photobleaching Fast (1 MHz) and Slow (250 kHz) Masking for ROIs Customize Laser Power at Each Slice Using Software Control
Variable Attenuator		Manual and Computer Control of Laser Power in Systems Without a Pockels Cell Improves Pockels Cell Performance One-Click Shutter
Variable Beam Expander		1X - 3X Beam Diameter Modulation into the Objective Back Aperture Using Software Control
Beam Stabilizer		Maintain Stable Beam Pointing During Laser Excitation, Laser Wavelength Switching, and Temporal Drift
Volume Imaging Using Bessel Beams		3D Video-Rate for Volumetric Functional Imaging Enhanced Temporal Resolution Adequate for Studying Internal Systems at Cellular Lateral Resolution In Vivo Press Release

Sample Holders
Rigid Stands for Slides, Recording Chambers, or Platforms		Minimal Footprint Conserves Space Around Objective and the Microscope Slim Profile Leaves Room for Dodt or DIC Imaging Modules Excellent Long-Term Stability Easily Rotate Samples Into and Out of the Beam Path
XY Platforms for Micromanipulators		Large Working Space that Surrounds the Objective on Three Sides Ideal for Setups Where the Sample and Apparatus Need to Move in Unison, Such as Patch Clamping 2" Travel in X and Y; 0.5 µm Encoder Resolution
Gibraltar Platform		Large, Stable Working Space for Sample and Supplementary Equipment Honeycomb Breadboard for Vibration Stability Open Design Allows for Unrestricted Instrument Operation

Thorlabs Support
Fully Designed and Manufactured In-House		Engineers Work Under One Roof to Lower Your Costs and Create Seamless Solutions Expertise in Every System Component
Modular System Construction		As Your Experimental Needs Evolve, Upgrade Your Microscope Without Sacrificing Existing Capabilities
Professional Installation		Thorlabs Technician Visits Your Lab to Assemble, Test, and Demonstrate Use of Your Microscope
Quick Support		Thorlabs Technicians and Application Specialists Available for Videoconferencing Communicate with Our Support Staff Faster than an Engineer Could Travel to Your Location Thorlabs Will Ship You a Camera with a Microphone to Facilitate the Conversation With Permission, Thorlabs Will Remote Desktop in to Address Software Issues

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Bergamo^® II Modules

Thorlabs Bergamo^® II microscopes are modular systems that can be customized in the design process to meet the exact needs of the experiment. The modules listed below are displayed in a variety of pre-built examples found on our Configurations tab to help provide a starting point for your design.

Galvo-Resonant Scanners, Galvo-Galvo Scanners, and Spatial Light Modulators

Bergamo^® II microscopes can be configured with one or two co-registered scan paths to propagate, condition, and direct an input laser beam. Each path can utilize a resonant-galvo-galvo scanner, galvo-resonant scanner, galvo-galvo scanner, and/or a spatial light modulator (SLM). These choices allow the user to optimize each experiment as needed for high frame rates, high sensitivity, and/or targeted exposure of the regions of interest.

Resonant-Galvo-Galvo Scanners for Random Access Scanning
Thorlabs offers 8 kHz and 12 kHz resonant-galvo-galvo (RGG) scanners. These scanners enable multiple areas within a single FOV to be imaged at high speed in quick succession. Our 8 kHz scanners utilize the entire field of view and offer a maximum frame rate of 400 fps, while our 12 kHz scanners provide an increased frame rate of 600 fps.

Galvo-Resonant Scanners for High-Speed Imaging
Thorlabs offers 8 kHz and 12 kHz galvo-resonant scanners. Our 8 kHz scanners utilize the entire field of view and offer a maximum frame rate of 400 fps, while our 12 kHz scanners provide an increased frame rate of 600 fps.

Galvo-Galvo Scanners for User-Defined ROI Shapes
Galvo-galvo scanners support user-drawn scan geometries (lines, polylines, squares, and rectangles) and also support custom photoactivation patterns (circles, ellipses, polygons, and points). They offer consistent pixel dwell times for better signal integration and image uniformity.

Spatial Light Modulator for Simultaneous Targeting
Unlike scanners, which physically move from point to point, spatial light modulators (SLMs) use holography to diffract the beam and shape it in a user-defined pattern. This includes general beam shaping as well as the creation of multiple focal points at the FOV; the latter allows multiple sites in a sample to be photoexcited simultaneously.

Scan Paths of Example Configurations

B243:	Multi-Target Photoactivation (Rotating)	Path 1: Galvo-Resonant Path 2: SLM
B242:	Two- and Three-Photon Imaging (Rotating)	Path 1: Galvo-Resonant Path 2: Galvo-Galvo
B251:	Random Access Scanning (Rotating)	Resonant-Galvo-Galvo
B241:	In Vivo Two-Photon Imaging (Rotating)	Galvo-Resonant
B252:	Dual-Path Random Access Scanning (XYZ)	Path 1: Resonant-Galvo-Galvo Path 2: Galvo-Galvo
B262:	Dual-Path Confocal Imaging (XYZ)	Path 1: Galvo-Resonant Path 2: Galvo-Galvo
B231:	Simple Imaging (XYZ)	Galvo-Resonant
B211:	Video and High-Speed Imaging (Z-Only)	Galvo-Galvo
B201:	Simple Imaging (Z-Only)	Galvo-Galvo

Figure 3. Wavelength Switching Using Tiberius Ti:Sapphire Laser Shown at 1/16^th Actual Speed

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Figure 2. Fast Switching between the optimal excitation wavelengths of 750 nm and 835 nm provides the high contrast seen in this composite image. The two-channel set was collected at an imaging rate of 7 fps.

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Figure 1. The above image was acquired using single-wavelength excitation at 788 nm, while the optimum excitation wavelengths for the two tags are 750 nm and 850 nm.

Fast Switching Using a
Tunable Femtosecond Laser

With an industry-leading tuning speed of up to 4000 nm/s and a wide 720 to 1060 nm tuning range, the Tiberius^® Ti:Sapphire Femtosecond Laser is ideal for fast sequential imaging in multiphoton microscopy applications.

A 25 µm thick sagittal section of an adult rat brain is shown in the images and video to the right. The red channel corresponds to fluorescence from chick anti-neurofilament that is optimally excited at 835 nm, while the green channel corresponds to fluorescence from mouse anti-GFAP that is optimally excited at 750 nm.

Figure 1 shows fluorescence from single-wavelength excitation at 788 nm, which sub-optimally excites the two tags simultaneously. Figure 2 is a composite image of the fluorescence produced by a two-color excitation image sequence acquired at 7 fps where the excitation wavelength was rapidly tuned between 750 and 835 nm. The video in Figure 3 shows the fast-switching used to create the composite image in Figure 2 at 1/16^th of the actual speed. When compared to single-wavelength excitation at 788 nm with the same intensity, fast switching offers much higher image contrast as it provides optimal excitation of both fluorophores.

This immunofluorescence sample was prepared by Lynne Holtzclaw of the NICHD Microscopy and Imaging Core Facility, a part of the National Institutes of Health (NIH) in Bethesda, MD.

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Figure 2. Close-Up of a Gaussian Beam

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Figure 1. Close-Up of a Bessel Beam

Volume Imaging Technique Using Bessel Beams

Thorlabs is excited to offer a new ultra-fast imaging technique that uses a bessel beam to provide video-rate volumetric functional imaging of neuronal pathways and interactions in vivo. These unique beams are non-diffractive and self-healing, which allows them to maintain a tight focus and even reform as they pass through tissue. This technique will be offered for Thorlabs' Bergamo II multiphoton microscopes and Thorlabs' Multiphoton Mesoscope.

The images to the right depict a bessel beam and a gaussian beam, respectively. As you can see in the images, the gaussian beam has a singular point of focus that progressively becomes weaker as it diverges from the central point, whereas the bessel beam has a beam annulus that maintains its focus.

To read the press release about this new technique, click here.

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Rotating Bergamo II systems are outfitted with multi-joint articulating periscopes. This periscope's design offers the enhanced flexibility needed to allow the entire scanning system to be tilted with respect to the sample.

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Upright Bergamo II systems are equipped with periscopes that permit the microscope's full travel range in X, Y, and Z to be used without compromising the optical performance.

Periscopes

Most lasers used in multiphoton microscopy are delivered by a free-space beam. The Bergamo II's ability to translate the objective around the focal plane in up to four axes (X, Y, Z, and θ) also requires the beam path to translate along the same axes while maintaining alignment. Bergamo II systems overcome this engineering challenge using multi-jointed periscopes.

Configurations with Articulating Periscopes

B243:	Multi-Target Photoactivation (Rotating)
B242:	Two- and Three-Photon Imaging (Rotating)
B251:	Random Access Scanning (Rotating)
B241:	In Vivo Two-Photon Imaging (Rotating)

Configurations with Fixed Periscopes

B252:	Dual-Path Random Access Scanning (XYZ)
B262:	Dual-Path Confocal Imaging (XYZ)
B231:	Simple Imaging (XYZ)
B211:	Video and High-Speed Imaging (Z-Only)
B201:	Simple Imaging (Z-Only)

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Emission filters and dichroic cubes are held behind magnetically sealed doors on the front of the PMT detection module.

Collection Optics of Example Configurations^a

B243:	Multi-Target Photoactivation (Rotating)	Epi: 10°
B242:	Two- and Three-Photon Imaging (Rotating)	Epi: 14°
B251:	Random Access Scanning (Rotating)	Epi: 14°
B241:	In Vivo Two-Photon Imaging (Rotating)	Epi: 14°
B252:	Dual-Path Random Access Scanning (XYZ)	Epi: 14° Transmitted: 13°
B262:	Dual-Path Confocal Imaging (XYZ)	Epi: 14°
B231:	Simple Imaging (XYZ)	Epi: 8° Transmitted: 13°
B211:	Video and High-Speed Imaging (Z-Only)	Epi: 8°
B201:	Simple Imaging (Z-Only)	Epi: 8°

Super Broadband Scan Optics

Bergamo II microscopes feature proprietary scan optics that are optimized and corrected for excitation wavelengths within the 450 - 1100 nm, 680 - 1600 nm, or 900 - 1900 nm wavelength range, ideal for photostimulation, two-photon imaging, and three-photon imaging, respectively. These broad ranges, extending from the visible well into the near infrared, were chosen to support the latest widely tunable Ti:Sapphire lasers and OPO systems, as well as dual-output lasers such as the Chameleon Discovery.

Our optics take full advantage of the optical designs used in the low-magnification, high-numerical-aperture objectives by filling the back aperture of the objective up to Ø20 mm. This creates an exceptional scan area that lets you find a region of interest more quickly or simply image more cells at once.

Large-Angle Signal Collection Optics

Deriving the most signal from limited photons is the fundamental goal of any detection system. By positioning the PMTs immediately after the objective (a "non-descanned" geometry), light that is scattered by the sample, which therefore appears to originate outside the objective's field of view, still strikes the PMTs and adds to the collected signal. This is a benefit unique to multiphoton microscopy. Collecting beyond the objective's design field of view greatly enhances overall detection efficiency when imaging deep in tissue.

In the epi direction, we offer signal collection angles^a of 8°, 10°, or 14°, while in the transmitted direction, we offer a signal collection angle^a of 13°. Our collection modules can optionally be outfitted with mechanical shutters for photoactivation experiments.

Easy-to-Reach Emission Filters and Dichroic Holders

Bergamo II systems are fully compatible with industry-standard fluorescence filter sets that include Ø25 mm fluorescence filters and 25 mm x 36 mm dichroic mirrors. Unlike competing designs, Thorlabs' detector modules have magnetic holders that make it simple and quick to exchange filters for different measurements.

We also offer detection modules for large-area Ø32 mm fluorescence filters and 32 mm x 42 mm dichroics, which support greater collection angles for increased signal.

Detectors in Epi and Transmitted Directions

We employ high-sensitivity GaAsP PMTs in our multiphoton systems, which can offer high quantum efficiency, aiding in imaging weakly fluorescent or highly photosensitive samples. Our PMTs can either be thermoelectrically cooled for improved sensitivity toward weak signals or non-cooled for a smaller package size and greater numerical aperture. Multialkali PMTs are also available.

All Bergamo^® II microscopes can be equipped with either two or four detection channels in the epi direction, and/or two detection channels in the forward direction. The user can configure the forward-direction channels to detect the same fluorescent tags as the epi-direction PMTs, raising the microscope's sensitivity toward thin, weakly fluorescent specimens.

A maximum of four channels can be controlled by the software at a given time.

Angles Quoted for an Objective with a Ø20 mm Entrance PupilXXX

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Multi-Axis Controller with Touchscreen

This controller is specifically designed for rotating Bergamo II microscope bodies. It uses knobs to control up to five motorized axes. On rotating systems, a rocker switch changes between fine objective focusing and translation of the elevator base. Each axis can be disabled on an individual basis in order to maintain a location along the desired direction.

The integrated touchscreen lets two spatial locations be saved and retrieved locally. Up to eight spatial locations can be saved on the computer running ThorImage^®LS. The touchscreen also reads out the position of every motor.

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Objectives

Bergamo II microscopes accept infinity-corrected objectives with M34 x 1.0, M32 x 0.75, M25 x 0.75, or RMS threads. Together, these options encompass the majority of low-magnification, high-NA objectives used in multiphoton microscopy. With a large field number of 20, our scan optics completely utilize the optical designs of these specialized objectives, offering enhanced light-gathering ability compared to competing microscopes using the same objectives.

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Rigid Stand Sample Holders

Thorlabs' Rigid Stands are rotatable, lockable, low-profile platforms for mounting slides, recording chambers, our Z-axis piezo stages, and custom experimental apparatuses. Each fixture is supported by a solid Ø1.5" stainless steel post for passive vibrational damping, which is in turn held to the workstation by the red post holder.

A locking collar maintains the height of the platform, allowing it to easily rotate into and out of the optical path, and a quick-release mechanism holds the post in place once the desired position is achieved.

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A Quantulux sCMOS Camera and 1.4 Megapixel Scientific Camera

Scientific Cameras

Our low-noise, scientific-grade CCD, sCMOS, and CMOS cameras were designed for full compatibility with Thorlabs’ multiphoton microscopy systems. Useful for widefield and fluorescence microscopy, they are capable of visualizing in vitro and in vivo samples using reflected light and fluorescence emission. They work in conjunction with the epi-fluorescence module to help locate fiducial markers, and they also enable imaging modalities that do not require laser exposure.

Thorlabs' cameras are driven by our internally developed ThorCam software package. CCD cameras are available in 1.4 MP, 4 MP, 8 MP, and fast-frame-rate versions, the sCMOS camera is available with a 2.1 MP sensor, and our CMOS cameras are available with a 5 MP sensor. Generally speaking, cameras with lower resolution offer higher maximum frame rates. These cameras also feature a separate auxiliary port that permits the image acquisition to be driven by an external electrical trigger signal.

Bergamo^® II microscopes are also directly compatible with any camera using industry-standard C-mount or CS-mount threads.

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Trans-Illumination Module, Motorized Condenser Stage, and
Rigid Stand Sample Holder Underneath the Objective

User-Installable Dodt Contrast and DIC Imaging Modules

The modular construction of the Bergamo^® II makes it exceptionally easy for the user to convert the microscope between in vitro and in vivo applications. Our user-installable trans-illumination modules for Dodt contrast, laser-scanned Dodt contrast, and differential interference contrast (DIC) take less than 5 minutes to attach or remove from the microscope body. These modules are available for both rotating and upright bodies.

Each option is paired with our basic 3-axis controller, which optimizes the illumination conditions by translating our motorized condenser stage over a 1" range. This versatile design is compatible with air and high-NA oil immersion condensers designed by Nikon.

To complement these modules, we manufacture slim-profile rigid stand sample holders that are ideal for positioning slides between the transmitted light module and the objective.

Configurations with Trans-Illumination Module

B252:	Dual-Path Random Access Scanning (XYZ)
B231:	Simple Imaging (XYZ)

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Structural Neurobiology

Neurological Disorders

Neural Development and Plasticity

Neurogenetics

Functional and Molecular Imaging

Synapses and Circuits

Ion Channels, Transporters, and Neurotransmitter Reporters

Cell Biology of Neurons, Muscles, and Glia

Drug Discovery

1.2 mm In Vivo Deep Brain 3D Image Stack, Courtesy of Dr. Hajime Hirase, Katsuya Ozawa, and YOU. H, RIKEN Brain Science Institute, Wako, Japan

Application Summary

Scientists investigate the structure of the brain to understand functions of neuronal proteins as well as the causes of neurological diseases. Due to the difficulty of imaging through brain tissue caused by light scattering, this study often requires a multi-modality configuration allowing for a range of experimental conditions using any combination of multiphoton, confocal, and epi-fluorescence imaging. The three example multiphoton microscope configurations in the table below are designed to accommodate the needs of Structural Biology. Each configuration features fast-Z power ramping to accomplish high-resolution imaging deep within a sample. Our Two- and Three-Photon Imaging configuration uses both galvo-resonant and galvo-galvo scanners and infrared wavelength scanning optics to image second- and third-harmonic generation (SHG and THG). Alternatively, our Dual-Path Multiphoton Microscope with Confocal Imaging is outfitted with a confocal path that accommodates up to 4 laser lines and a 4-channel PMT detection module. The addition of a six-cube epi-illuminator module and sCMOS Quanatlux camera allows this system to perform epi-fluorescence imaging. Our Simple XYZ Imaging configuration is well-balanced for both in vitro and fixed stage in vivo microscopy research. With a removable transmitted illumination module, this versatile system can support a wide variety of experimental techniques, imaging modalities, and sample subjects.

Body	Rotating	Upright XYZ
Function	Two- and Three-Photon Imaging	Dual Path with Confocal Imaging	Simple XYZ Imaging
Configuration	B242	B262	B231

Publications

Lee KS, Huang X, and Fitzpatrick D. "Topology of ON and OFF inputs in visual cortex enables an invariant columnar architecture." Nature. 2016 May 5; 533: 90-94.

Lang X and Lyubovitsky JG. "Structural dependency of collagen fibers on ion types revealed by in situ second harmonic generation (SHG) imaging Ion channels and G coupled receptors." Analytical Methods. 2014 Nov 13; 7 (5): 1637-2230.

Ehmke T, Nitzsche TH, Knebl A, and Heisterkamp A. "Molecular orientation sensitive second harmonic microscopy by radially and azimuthally polarized light." Biomedical Optics Express. 2014 Jul 1; 5 (7): 2231-2246.

Lang X, Spousta M, Hwang YJ, and Lyubovitsky JG. "Noninvasive imaging of embryonic stem cell cultures by multiphoton microscopy reveals the significance of collagen hydrogel preparation parameters." Analytical Methods. 2011 Jan 20; 3 (3): 529-536.

Cai F, Yu J, Qian J, Wang Y, Chen Z, Huang J, Ye Z, and He S. "Use of tunable second-harmonic signal from KNbO₃ nanoneedles to find optimal wavelength for deep-tissue imaging." Laser & Photonics Review. 2014 Jun 17; 8 (6): 865-874.

Poguzhelskaya E, Artamonov D, Bolshakova A, Vlasova O, and Bezprozvanny I. "Simplified method to perform CLARITY imaging." Molecular Neurodegeneration. 2014 May 26; 9 (19).

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Stitched Confocal Fluorescence Image of Rat Retina Stained with DAPI, Alexa Fluor^® 555 and Alexa Fluor^® 633, Courtesy of Dr. Jennifer Kielczewski, National Eye Institute, National Institutes of Health, Bethesda, MD

Application Summary

Researching neurological disorders involves measuring neural function using two-photon calcium imaging. This research requires fast image acquisition and photostimulation. We recommend three of our multiphoton microscope configurations for this application (see the table below). Each configuration offers a large working space and rotating microscope body, making it ideal for in vivo animal studies. Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows the activation of groups of neurons at varying depths within a single field of view. For increased penetration of samples with scattering tissues, the three-photon capability of our Two- and Three-Photon Imaging configuration is recommended. Our Random-Access Scanning configuration uses a resonant-galvo-galvo scanner to take multiple high-resolution images within a single field of view. This scanner provides all the speed of a resonant-galvo scanner, while enabling multiple user-defined fluorescence activation regions for correlating neural responses in multiple regions of the brain.

Body	Rotating
Function	Multi-Target Photoactivation	Two- and Three-Photon Imaging	Random Access Scanning
Configuration	B243	B242	B251

Publications

Murphy SC, Palmer LM, Nyffeler T, Müri RM, and Larkum ME. "Transcranial magnetic stimulation (TMS) inhibits cortical dendrites." eLife. 2016 Mar 18; 5: e13598.

Lalchandani RR, van der Goes MS, Partridge JG, and Vicini S. "Dopamine D₂ receptors regulate collateral inhibition between striatal medium spiny neurons." The Journal of Neuroscience. 2013 Aug 28; 33 (35): 14075-14086.

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Two-Photon Image of Neurons Expressing Thy1-YFP in a Cleared Region of the Hippocampus, Courtesy of the 2017 Imaging Structure and Function in the Nervous System Course at Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

Application Summary

Scientists interested in this application focus on tracing neuronal pathways and researching dendritic spine plasticity. Imaging systems used in this type of research are capable of two-photon calcium imaging and/or confocal fluorescence imaging. High-resolution and high-sensitivity are crucial features for these systems. We offer three multiphoton configurations that are ideal for this application (see the table below). Each configuration may be used in photon-limited environments as they feature sensitive detection modules, in addition to our 10° or 14° full field-of-view collection optics with ultrasensitive GaAsP PMTs. Our Dual-Path with Confocal Imaging configuration allows for a range of experimental conditions to be observed using any combination of multiphoton, confocal, and epi-fluorescence imaging, along with photoactivation. Alternatively, the Video and High-Speed Imaging and Simple Z-Axis Imaging configurations provide a small footprint and large throat depth for in vivo two-photon imaging.

Body	Upright XYZ	Upright Z-Axis
Function	Dual Path with Confocal Imaging	Video and High-Speed Imaging	Simple Z-Axis Imaging
Configuration	B262	B211	B201

Publications

Gillet SN, Kato HK, Justen MA, Lai M, and Isaacson JS. "Fear learning regulates cortical sensory representations by suppressing habituation." Frontiers in Neural Circuits. 2018 Jan 10; 11: 112.

Micu I, Brideau C, Lu L, and Stys PK. "Effects of laser polarization on responses of the fluorescent Ca²⁺ indicator X-Rhod-1 in neurons and myelin." Neurophotonics. 2017 Jun 5; 4 (2): 025002.

Chen SX, Kim AN, Peters AJ, and Komiyama T. "Subtype-specific plasticity of inhibitory circuits in motor cortex during motor learning." Nature Neuroscience. 2015 Jun 22; 18: 1109-1115.

Peters AJ, Chen SX, and Komiyama T. "Emergence of reproducible spatiotemporal activity during motor learning." Nature. 2014 Jun 12; 510: 263-267.

Laser-Scanned Two-Photon SHG+Dodt Gradient Contrast of Zebrafish Embryo

Application Summary

Neurogenetic studies require a wide variety of experimental setups for in vivo research. Multiphoton imaging is ideal for studying live organisms, especially embryos, as it reduces the occurrence of photobleaching and phototoxicity that is common with other light microscopy techniques. There are three multiphoton configurations we suggest for Neurogenetic applications (see the table below). The small footprint and large throat depth of each system provide ample room for sample mounts and experimental apparatus, such as the large setup used for Drosophilia studies by Chen et al. (click here for supplementary videos). Additionally, these systems provide video-rate, sequential two-photon imaging to study fast dynamic biological and chemical processes in vivo without damaging the sample.

Body	Upright XYZ	Upright Z-Axis
Function	Simple XYZ Imaging	Video and High-Speed Imaging	Simple Z-Axis Imaging
Configuration	B231	B211	B201

Publications

Chen CL, Hermans L, Viswanathan MC, Fortun D, Aymanns F, Unser M, Cammarato A, Dickinson MH, and Ramdya P. "Imaging neural activity in the ventral nerve cordof behaving adult Drosophila." Nature Communications. 2018 October 22; 9: 4390.

Dechen K, Richards CD, Lye JC, Hwang JEC, and Burke R. "Compartmentalized zinc deficiency and toxicities caused by ZnT and Zip gene over expression result in specific phenotypes in Drosophila." The International Journal of Biochemistry & Cell Biology. 2015 Jan 3; 60: 23-33.

Imaging Visually Evoked Synaptic Calcium Transient In Vivo, Using Dendrite Labels with GCAMP6, 200 µm Deep in Layer 2/3 Visual Cortex, Courtesy of David Fitzpatrick, Max Plank Institute for Neurobiology, Jupiter, FL, USA

Application Summary

In vivo functional imaging of organisms and the individual neurons linked to specific organism behaviors requires high-speed and high-sensitivity imaging. We offer six configurations for this application (see the table below). These configurations are equipped with an 8 or 12 kHz galvo-resonant imaging scanner and our 10° or 14° wide-angle collection optics to enable fast, high-resolution imaging. Each microscope system features a large throat depth, 5” of vertical travel, and smooth movement along the Z-axis to create a large working space ideal for in vivo volume imaging deep into highly scattering samples of neural tissue. For an improved range of movement around a sample, we recommend a configuration with a rotating body.

Body	Rotating			Upright XYZ		Upright Z-Axis
Function	Multi-Target Photoactivation	Random Access Scanning	In Vivo Two-Photon Imaging	Dual-Path with Confocal Imaging	Simple XYZ Imaging	Video and High-Speed Imaging
Configuration	B243	B251	B241	B262	B231	B211

Publications

Aghayee S, Bowen Z, Winkowski DE, Harrington M, Kanold P, and Losert W. "Particle tracking facilitates real time motion compensation in 2D or 3D two-photon imaging of neuronal activity." Frontiers in Neural Circuits. 2017 Aug 15; 11: 56.

Schnell B, Ros IG, and Dickinson MH. "A descending neuron correlated with the rapid steering maneuvers of flying Drosophila." Current Biology. 2017 Apr 6; 27 (8): 1200-1205.

Roth MM, Dahmen JC, Muir DR, Imhof F, Martini FJ, and Hofer SB. "Thalamic nuclei convey diverse contextual information to layer 1 of visual cortex." Nature Neuroscience. 2015 Dec 21; 19: 299-307.

Barnstedt O, Keating P, Weissenberger Y, King AJ, and Dahmen JC. "Functional microarchitecture of the mouse dorsal inferior colliculus revealed through in vivo two-photon calcium imaging." The Journal of Neuroscience. 2015 Aug 5; 35 (31): 10927-10939.

Boyd AM, Kato HK, Komiyama T, and Isaacson JS. "Broadcasting of cortical activity to the olfactory bulb." Cell Reports. 2015 Feb 24; 10 (7): 1032-1039.

Simultaneous Photostimulation of 100 Cells Co-Expressing GCaMP6f (Green) and C1V1 (Red), Courtesy of Lloyd Russell, Dr. Adam Packer, and Professor Michael Häusser, University College London, United Kingdom

Application Summary

By studying synapses and circuits, researchers are able to understand neuronal activity. Imaging synapses and circuits often requires simultaneous stimulation of populations of neurons. To achieve this, we offer three configurations of our multiphoton microscope capable of fast image acquisition of multiple regions within a single field of view (see table below). Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows multiple sites in a sample to be photoexcited simultaneously. With the SLM, each beamlet can be shaped to improve the efficacy of photoactivation, a crucial feature for activating neural populations at varying depths within a single field of view (FOV). Our High-Speed, Random-Access Scanning configuration uses a resonant-galvo-galvo scanner to take multiple high-resolution images within a single field of view. This scanner provides all the speed of a resonant-galvo scanner, while enabling multiple user-defined photoactivation regions. Lastly, our In Vivo Two-Photon Imaging configuration uses a galvo-resonant scanner for high-speed imaging.

Body	Rotating
Function	Multi-Target Photoactivation	Random Access Scanning	In Vivo Two-Photon Imaging
Configuration	B243	B251	B241

Publications

Scholl B, Wilson DE, and Fitzpatrick D. "Local order within global disorder: synaptic architecture of visual space." Neuron. 2017 Dec 6; 96 (5): 1127-1138.

Leinweber M, Ward DR, Sobczak JM, Attinger A, and Keller GB. "A sensorimotor circuit in mouse cortex for visual flow predictions." Neuron. 2017 Sep 13; 95 (6): 1420-1432.

Iacaruso, MF, Gasler IT, and Hofer SB. "Synaptic organization of visual space in primary visual cortex." Nature. 2017 Jul 27; 547: 449-452.

Dipoppa M, Ranson A, Krumin M, Pachitariu M, Carandini M, and Harris KD. "Vision and locomotion shape the interactions between neuron types in mouse visual cortex." Neuron. 2016 May 2; 98 (3): 602-615.

Cochlear Organotypic Culture Loaded with Fluo4-AM and DM-Nitrophen AM, Calcium Release Triggered Using Galvo-Galvo Uncaging Pathway in Outer Hair Cell Indicated by Red Box After ~13 Seconds, Courtesy of Federico Ceriani and Walter Marcotti, University of Sheffield

Application Summary

In this application, scientists are interested in neural connections and intercellular movement. With multiphoton imaging, they are able to trace the direction and speed of ions moving through channel membrane proteins or neurotransmitters moving from one neuron to another. This research requires a microscopy system that has high-resolution and high-speed imaging of multiple fields of view (FOVs). We recommend six of our multiphoton configurations for this application (see the table below). This area of research often requires both in vivo and in vitro imaging within the same study, so within each configuration, our transmitted light modules can be installed or removed by the user in just a few minutes, making it exceptionally easy to switch between the two imaging modalities. These configurations are capable of high-frame-rate imaging and targeted laser activation for photostimulation, making them ideal for correlating neural responses in multiple regions of the brain.

Body	Rotating			Upright XYZ		Upright Z-Axis
Function	Multi-Target Photoactivation	Random Access Scanning	In Vivo Two-Photon Imaging	Dual-Path Random Access Scanning	Dual-Path with Confocal Imaging	Video and High-Speed Imaging
Configuration	B243	B251	B241	B252	B262	B211

Publications

De Toni L, Garolla A, Menegazzo M, Magagna S, Di Nisio A, Šabovic I, Rocca MS, Scattolini V, Filippi A, and Foresta C. "Heat sensing receptor TRPV1 is a mediator of thermotaxis in human spermatozoa." PLoS One. 2016 Dec 16; 11 (12): e0167622.

Mongeon R, Venkatachalam V, and Yellen G. "Cytosolic NADH-NAD⁺ redox visualized in brain slices by two-photon fluorescence biosensor imaging." Antioxidants & Redox Signaling. 2016 Oct 1; 25 (10): 553-563.

Lewin AE, Vicini S, Richardson J, Dretchen KL, Gillis RA, and Sahibzada N. "Optogenetic and pharmacological evidence that somatostatin-GABA neurons are important regulators of parasympathetic outflow to the stomach." The Journal of Physiology. 2016 Mar 9; 594 (10): 2661-2679.

Cossell L, Iacaruso MF, Muir DR, Houlton R, Sader EN, Ko H, Hofer SB, and Mrsic-Flogel TD. "Functional organization of excitatory synaptic strength in primary visual cortex." Nature. 2015 Feb 19; 518: 399-403.

Click for Full Size Image

Top: Astrocytes are labelled with SR101 (red). Arrows point to astrocytes that had Ca²⁺ elevations during tDCS. The numbers correspond to the cells and neurogliopil regions plotted in the graphs below. Bottom: Fluorescent intensity (ΔF/F) traces of astrocytes (orange), neurons (green) and neurogliopil (brown). Figure Courtesy of Monai H. et al. (See Below)

Application Summary

The study of cell biology, muscles, and glia often involves imaging in vitro or in vivo samples tagged with multiple fluorophores. Through multiphoton imaging with these fluorescent markers, researchers are able to observe gene expression related to neural function in different areas of the brain. We recommend two of our configurations for this application, see the table below. With two to four channel detection modules and fast sequential imaging using our Tiberius^® Tunable fs laser, both of these configurations offer the flexibility necessary for experiments in this field. In addition, each configuration has a small footprint and a large throat depth to provide ample room for numerous sample mounting options, including in vivo imaging of mammalian brains via transcranial windows.

Body	Upright XYZ	Upright Z-Axis
Function	Simple XYZ Imaging	Video and High-Speed Imaging
Configuration	B231	B211

Publications

Rose T, Jaepel J, Hübener M, and Bonhoeffer T. "Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortex." Science. 2016 Jun 10; 352 (6291): 1319-1322.

Monai H, Ohkura M, Tanaka M, Oe Y, Konno A, Hirai H, Mikoshiba K, Itohara S, Nakai J, Iwai Y, and Hirase H. "Calcium imaging reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain." Nature Communications. 2016 March 22; 7: 11100.

Lu W, Xie Z, Tang Y, Bai L, Yao Y, Fu C, and Ma G. "Photoluminescent mesoporous silicon nanoparticles with siccr2 improve the effects of mesenchymal stromal cell transplantation after acute myocardial infarction." Theranostics. 2015 Jun 25; 5 (10): 1068-1082.

Qin X, Qiu C, and Zhao L. "Maslinic acid protects vascular smooth muscle cells from oxidative stress through Akt/Nrf₂/HO-₁ pathway." Molecular and Cellular Biochemistry. 2014 Feb 20; 390 (1-2): 61-67.

Click for Full Size Image

Artistic Rendering of DNA

Application Summary

Drug discovery research is rapidly expanding and often requires a wide variety of experimental setups and imaging techniques. Two-photon imaging is frequently used to measure the characteristics of drug applications, including depth of drug penetration and the area of its spread throughout the cortex. We offer two configurations that are suitable for this application, see the table below. These configurations are are well-balanced for both in vitro and fixed stage in vivo microscopy research. The modularity of the Bergamo systems' removable trans-illumination module provides versatility in regard to experimental techniques, imaging modalities, and sample subjects. The Gibraltar breadboard platform line is ideal for mounting samples and supplementary equipment within these configurations.

Body	Upright XYZ
Function	Simple XYZ Imaging	Dual-Path Random Access Scanning
Configuration	B231	B252

Publications

Strobl MJ, Freeman D, Patel J, Poulsen R, Wendler CC, Rivkees SA, and Coleman E. " Opposing effects of maternal hypo- and hyperthyroidism on the stability of thalamocortical synapses in the visual cortex of adult offspring." Cerebral Cortex. 2017 May 1; 27 (5): 3015-3027.

Scattolini V, Luni C, Zambon A, Galvanin S, Gagliano O, Ciubotaru CD, Avogaro A, Mammano F, Elvassore N, and Fadini GP. "Simvastatin rapidly and reversibly inhibits insulin secretion in intact single-islet cultures." Diabetes Therapy. 2016 Nov 9; 7 (4): 679-693.

Example Configurations

Explore the details of example Bergamo II rotating, XYZ, and Z-axis system configurations by clicking on the expandable sections below. The modular nature of our multiphoton microscopy platform allows us to modify configurations to meet individual experimental needs or adjust the functionality of a microscope after installation; more information on modules featured in the systems below can be found on the Modules tab.

Configuration

System Highlights

Rotating Body

Configuration B243:
Multi-Target Photoactivation

Click for Details

Key Features

Suggested Applications

Spatial Light Modulator (SLM) in Secondary Beam Path, Providing Holographic, All-Optical Multi-Target Stimulation
Large Throat Depth for In Vivo Animal Studies
Separate Lasers for Multiphoton Imaging and Photoactivation
Galvo-Resonant Scanning for High-Speed Image Acquisition

Synapses and Circuits
Ion Channels, Transporters, and Neurotransmitter Reporters
Functional and Molecular Imaging
Neurological Disorders

Additional Specifications for Multi-Target Photoactivation Configuration

Configuration Specifications
Highlighted System Specifications
FOV	12 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels
Photoactivation	Targeted: Holographic, Simultaneous Multi-Site Full Field: During Scanner Flyback
Microscope Components
Microscope Body	-45 to +45° Rotation
Primary Scan Path (Imaging Scanners)	Galvo-Resonant (8 or 12 kHz)
Secondary Scan Path	Spatial Light Modulator
Scan Path Wavelength Range	450 - 1100 nm or 680 - 1600 nm
Pockels Cell	Slow (250 kHz) on Primary Path
Variable Attenuator	Motorized
Multiphoton Detection	2 Non-Cooled GaAsP PMTs
Collection Optics Module	10° with Shutter
Objective Holder	Single
Objective	Nikon 16X (with Piezo Objective Scanner)
Sample Stage	None
Widefield Imaging	Single Cube Epi-Illuminator Module Single-Channel Mounted LED 4 MP CCD Scientific Camera with GigE Interface
Laser	Multiphoton Imaging: Tiberius^® Tunable fs Laser Photoactivation: Menlo Systems' BlueCut fs Fiber Laser

Our spatial light modulator (SLM) manipulates the phase of the stimulation laser beam to generate hundreds of user-determined focal points. The SLM phase mask pattern can be rapidly switched, enabling multiple individual focal points to be targeted independently of each other. Each beamlet can be shaped to improve the efficacy of photoactivation; a crucial feature for activating neural populations at varying depths within a single FOV.

This configuration utilizes two separate lasers for imaging and photostimulation; in addition to the highly configurable nature of the Bergamo microscope system, the scanning paths can be adjusted for a dual-output laser source.

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The SLM is installed on the secondary scan path, enabling photostimulation simultaneous with multiphoton imaging.

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The Tiberius fs tunable laser is used for multiphoton imaging, while Menlo Systems' BlueCut fiber laser is used for photostimulation.

Configuration B242:
Two- and Three-Photon Imaging

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Key Features

Suggested Applications

Dual-Channel Paths for Simultaneous 2P and 3P Imaging, or 3P Imaging with Photoactivation
Beam Conditioning with Variable Beam Expander, Pockels Cell, and Variable Attenuator
Simultaneous 2P or 3P Imaging with Widefield
Epi-Fluorescence
Large Working Space and Rotating Microscope Body for In Vivo Large Animal Studies

Structural Neurobiology
Neurological Disorders

Additional Specifications for 2P / 3P Imaging Configuration

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	Galvo-Resonant Scanner: 8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels Galvo-Galvo Scanner: 3 fps at 512 x 512 Pixels
Photoactivation	Targeted: Simultaneous IR, Sequential Visible
Microscope Components
Microscope Body	-5 to +95° Rotation
Primany Scan Path	Galvo-Resonant (8 or 12 kHz)
Secondary Scan Path	Galvo-Galvo (Ø4 mm or Ø5 mm)
Scan Path Wavelength Range	680 - 1600 nm on Primary Path 900 - 1900 nm on Secondary Path
Variable Beam Expander	Primary Path
Pockels Cell	Fast (1 MHz) on Primary Path Slow (250 kHz) on Secondary Path
Variable Attenuator	Motorized
Beam Stabilization	Primary and Secondary Paths
Multiphoton Detection	2 Cooled GaAsP PMTs
Collection Optics Module	14° with Shutter
Objective Holder	Single
Objective	Nikon 25X (with Piezo Objective Scanner)
Sample Stage	None
Widefield Imaging	Single-Cube Epi-Illuminator Module Four-Channel LED Source Quantalux™ sCMOS Scientific Camera
Laser	Primary Path: 3P Pulsed Laser Secondary Path: Tiberius^® fs Tunable Laser

This dual-path configuration uses both galvo-resonant and galvo-galvo scanners and infrared wavelength scanning optics to image second- and third-harmonic generation (SHG and THG).

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The primary and secondary scan path optics are optimized for 2P and 3P imaging, respectively. The beam paths are co-registered for simultaneous multi-channel imaging.

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View your subject at angles up to +95° with our Rotating Microscope Bases. The body rotation is centered on the focal point of your objective, allowing free movement without the need to refocus.

Configuration B251:
Random Access Scanning

Resonant-Galvo-Galvo Scanning Configuration

Click for Details

Key Features

Suggested Applications

Resonant-Galvo-Galvo Scanner for High-Resolution, Fast Acquisition of Multiple Regions within a Single FOV
Integrate with Soundbox for Sound-Sensitive Experiments
Edge Blanking with Fast Pockels Cell
Simultaneous Multi-Channel Epi-Fluorescence
Tiberius fs Ti:Sapphire Tunable Laser

Synapses and Circuits
Ion Channels, Transporters, and Neurotransmitter Reporters
Functional and Molecular Imaging
Neurological Disorders

Additional Specifications for Random Access Scanning Configuration

This high-speed random access scanning configuration uses a resonant-galvo-galvo scanner to take multiple high-resolution images within a single field of view. This scanner provides all the speed of a resonant-galvo scanner while enabling multiple user-defined regions to be chosen. This imaging functionality is ideal for correlating neural responses in multiple regions of the brain.

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels
Photoactivation	Targeted: Sequential Full Field: During Scanner Flyback
Microscope Components
Microscope Body	-5 to +95° Rotation
Imaging Scanners	Resonant-Galvo-Galvo (8 or 12 kHz)
Scan Path Wavelength Range	450 - 1100 nm
Pockels Cell	Fast (1 MHz)
Variable Attenuator	Motorized
Multiphoton Detection	2 Non-Cooled GaAsP PMTs
Collection Optics Module	14°
Objective Holder	Single
Objective	Olympus 20X (with Piezo Objective Scanner)
Sample Stage	Rigid Stand with Breadboard Insert
Widefield Imaging	Single-Cube Epi-Illuminator Module Single-Channel Mounted LED Quantalux™ sCMOS Scientific Camera
Laser	Tiberius^® Tunable fs Laser

RGG Random Access Scanning Configuration with Soundbox

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The microscope can be placed within a soundbox for sensitive auditory studies, as shown here.

Configuration B241:
In Vivo Two-Photon Imaging

Click for Details

Key Features

Suggested Applications

Large Working Space with Fully Rotatable Microscope
Integrate with Soundbox for Sound-Sensitive Experiments
Galvo-Resonant Scanner for High-Speed Imaging
Edge Blanking with Fast or Standard Pockels Cell
Simultaneous Multi-Channel Epi-Fluorescence
Tiberius^® fs Ti:Sapphire Tunable Laser

Ion Channels, Transporters, and Neurotransmitter Reporters
Functional and Molecular Imaging
Synapses and Circuits
Auditory Functional Imaging

Additional Specifications for In Vivo 2P Imaging Configuration

This rotating Bergamo microscope configuration is compact enough to be placed in an enclosure, enabling sound or light-sensitive in vivo studies to be performed.

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels
Photoactivation	Fixed Spot at the Center of the Field of View
Microscope Components
Microscope Body	-5 to +95° Rotation
Imaging Scanners	Galvo-Resonant (8 kHz or 12 kHz)
Scan Path Wavelength Range	450 - 1100 nm or 680 - 1600 nm
Pockels Cell	Fast (1 MHz) or Slow (250 kHz)
Variable Attenuator	Motorized
Multiphoton Detection	2 Non-Cooled GaAsP PMTs
Collection Optics Module	14° with Shutter
Objective Holder	Single
Objective	Olympus 20X (with Piezo Objective Scanner)
Sample Stage	Rigid Stand with Breadboard Insert
Widefield Imaging	Six-Filter-Turret Epi-Illuminator Module Solis™ High-Powered White LED Quantalux™ sCMOS Scientific Camera
Laser	Tiberius^® fs Tunable Laser

2P Rotating Microscope Encased in Soundbox

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2P Rotating Microscope Encased Inside a Soundbox

Upright XYZ Body

Configuration B252:
Dual-Path Random Access Scanning

Click for Details

Key Features

Suggested Applications

Light-Tight Enclosure for Laser Conditioning Modules and Components
Resonant-Galvo-Galvo Scanner for High-Speed, High-Resolution Imaging of Multiple FOVs
PMT Options for Scanned or De-Scanned Multiphoton Detection
Gibraltar Breadboard Platform for Samples and Supplementary Equipment
Dual-Output Spectra Physics InSight Laser
High-Speed, High-Resolution Volumetric Imaging Using Bessel Beams (Optional)

Ion Channels, Transporters, and Neurotransmitter Reporters
Drug Discovery
Ex Vivo Neurobiological Studies
Electrophysiology and Patch-Clamp Recordings
In Vivo Neurobiological Studies using Bessel Beam Volumetric Imaging Technique (Optional)

Additional Specifications for Dual-Path Random Access Scanning Configuration

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	Resonant-Galvo-Galvo Scanner: 8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels Galvo-Galvo Scanner: 3 fps at 512 x 512 Pixels
Scan Resolution	Bi-Directional: 8 kHz RGG and GG: Up to 2048 x 2048 Pixels 12 kHz RGG: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz RGG and GG: Up to 4096 x 4096 Pixels 12 kHz RGG: Up to 2336 x 2336 Pixels
Photoactivation	Targeted: Simultaneous IR, Sequential Visible Full Field: During Scanner Flyback
Microscope Components
Microscope Body	XYZ Motion
Primary Scan Path	Resonant-Galvo-Galvo (8 or 12 kHz)
Secondary Scan Path	Galvo-Galvo (Ø4 mm or Ø5 mm)
Scan Path Wavelength Range	Primary Path: 680 - 1600 nm Secondary Path: 450 - 1100 nm
Pockels Cell	Slow (250 kHz) on Primary Path
Variable Attenuator	Motorized
Multiphoton Detection	2 Cooled GaAsP PMTs 2 Non-Cooled GaAsP PMTs
Collection Optics Module	14° with Shutter
Objective Holder	Single
Objective	Nikon 25X (with Piezo Objective Scanner)
Sample Stage	Gibraltar Platform
Widefield Imaging	Transmitted Light Module with Laser Scanning Dodt Six-Filter-Turret Epi-Illuminator Module Fiber-Coupled 4-Channel LED Quantalux™ sCMOS Scientific Camera
Laser	Spectra Physics InSight Tunable Laser

Fully equipped for random access scanning, this configuration features both a resonant-galvo-galvo scanner on the primary path and a galvo-galvo scanner on the secondary path. The system uses a dual-output laser for multiphoton imaging.

Click for Details
The large Gibraltar stage shown above allows both samples and supplementary equipment to be aligned in the focal plane of the objective. Here, a motorized micromanipulator is mounted onto the breadboard to manipulate cells for patch-clamp recordings.

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A custom enclosure can be added to house beam conditioning modules and other components, creating a light-tight region in which parts may be altered without affecting the rest of the beam path.

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The resonant-galvo-galvo scanner on the primary path allows multiple regions of interest to be quickly scanned in a single field of view, while the galvo-galvo scanner on the secondary path can be used for simultaneous photoactivation.

Configuration B262:
Dual-Path with
Confocal Imaging

Click for Details

Key Features

Suggested Applications

Dual-Paths for Multiphoton Imaging with
Confocal Imaging or Photoactivation
Galvo-Resonant Scanner for High-Speed Imaging and Galvo-Galvo Scanner for Custom Geometric Scans
Four-Channel Confocal Fiber Laser and Tiberius^® fs Ti:Sapphire Tunable Laser

Structural Neurobiology
Ion Channels, Transporters, and Neurotransmitter Reporters
Neural Development and Plasticity
Functional and Molecular Imaging

Additional Specifications for Dual-Path Confocal Configuration

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	Galvo-Resonant Scanner: 8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels Galvo-Galvo Scanner: 3 fps at 512 x 512 Pixels
Scan Resolution	Bi-Directional: 8 kHz Galvo-Resonant and Galvo-Galvo: Up to 2048 x 2048 Pixels 12 kHz Galvo-Resonant: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Galvo-Resonant and Galvo-Galvo: Up to 4096 x 4096 Pixels 12 kHz Galvo-Resonant: Up to 2336 x 2336 Pixels
Photoactivation	Targeted: Simultaneous IR, Sequential Visible Full Field: During Scanner Flyback
Microscope Components
Microscope Body	XYZ Motion
Primary Scan Path (Multiphoton)	Galvo-Resonant (8 or 12 kHz)
Secondary Scan Path	Galvo-Galvo (Ø4 mm or Ø5 mm)
Scan Path Wavelength Range	450 - 1100 nm or 680 - 1600 nm on Primary and/or Secondary Path
Pockels Cell	Slow (250 kHz) (High-Speed and Wide-Wavelength-Range Options Available)
Variable Attenuator	Motorized
Multiphoton Detection	2 Non-Cooled GaAsP PMTs
Confocal Detection	4-Channel Multialkali PMTs with Variable-Size Pinhole Wheel
Collection Optics Module	14°
Objective Holder	Single
Objective	Nikon 25X (with Piezo Objective Scanner)
Sample Stage	Rigid Stand with Breadboard Insert
Widefield Imaging	Six-Filter-Turret Epi-Illuminator Module Broadband Light Source Quantalux™ sCMOS Scientific Camera
Laser	Multiphoton: Tiberius^® fs Tunable Laser Confocal: 4-Channel Viper Laser Source

This multi-path, multi-modality configuration allows for a range of experimental conditions to be observed using any combination of multiphoton, confocal, and epi-fluorescence imaging, along with photoactivation. Multiphoton imaging with photoactivation is enabled by two beam paths; the primary with a galvo-resonant scanner for high-speed imaging, and the secondary with a galvo-galvo scanner for area-specific photoactivation. The four-channel confocal laser connects through a fiber bulkhead to the secondary scan path, and the four-channel PMT detection module uses an additional port at the back focal plane of the objective. Epi-fluorescence is enabled by a six-filter-turret epi-illuminator module and sCMOS Quantalux camera.

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Thorlabs provides a variety of sample mounting options. The breadboard insert on the rigid stand provides the space and stability for a VR fly theater and supplementary cameras to be mounted below the objective, as shown.

Configuration B231:
Simple XYZ Imaging

Click for Details

Key Features

Suggested Applications

High-Speed Galvo-Resonant Scanning
High-Sensitivity GaAsP PMT
Free-Space Photodetector
Removable Transmitted Light Module with Laser Scanning Dodt Functionality
Epi-Fluorescence with Quantalux™ sCMOS Camera
Tiberius fs Ti:Sapphire Tunable Laser

Structural Neurobiology
Functional and Molecular Imaging
Drug Discovery
Fixed Stage Experiments
Neurogenetics
Cell Biology of Neurons, Muscles, and Glia

Additional Specifications for Simple XYZ Configuration

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	8 kHz Scanner: 30 fps at 512 x 512 Pixels or 12 kHz Scanner: 45 fps at 512 x 512 Pixels
Imaging Resolution	Bi-Directional: 8 kHz Scanner: Up to 2048 x 2048 Pixels 12 kHz Scanner: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Scanner: Up to 4096 x 4096 Pixels 12 kHz Scanner: Up to 2336 x 2336 Pixels
Photoactivation	Full Field: 4-Channel
Microscope Components
Microscope Body	XYZ Motion
Imaging Scanners	Galvo-Resonant (8 or 12 kHz)
Scan Path Wavelength Range	450 - 1100 nm
Pockels Cell	Slow (250 kHz) (High-Speed and Wide-Wavelength-Range Options Available)
Variable Attenuator	Motorized
Detection	2 Non-Cooled GaAsP PMTs
Collection Optics Module	8° with Shutter
Objective Holder	Manual Dual
Objective	Nikon 16X (with Piezo Objective Scanner) Nikon 40X
Sample Stage	Rigid Stand Slide Holder with Translation Stage
Widefield Imaging	Transmitted Light Module with Laser Scanning Dodt Single-Cube Epi-Illuminator Module Four-Channel LED Source Quantalux™ sCMOS Scientific Camera
Laser	Tiberius^® fs Tunable Laser

This configuration is well-balanced for both in vitro and fixed stage in vivo microscopy research. The modular system with a removable trans-illumination module provides high versatility for multiple experimental techniques, imaging modalities, and sample subjects.

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The ample space under the microscope enables large setups with supplemental equipment to be easily arranged under the objective. Here, we show a Drosophila VR theater stage along with two additional cameras. The setup can be synchronized with image acquisition using ThorSync, an add-on to our ThorImageLS software.

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Any one of our PMTs can be replaced with a free-space photodetector for signal detection of other imaging modalities. This image shows a variation of the microscope where one PMT is replaced with the DET10A2 for reflected signal detection.

Upright Z-Axis Body

Configuration B211:
Video and High-Speed Imaging

Click for Details

Key Features

Suggested Applications

High-Speed Galvo-Resonant Scanning
Pockels Cell and Motorized Variable Attenuator for Remote Laser Adjustment
2-Channel PMT Detection
Small Footprint with Large Throat Depth
Tiberius fs Ti:Sapphire Tunable Laser

Neural Development and Plasticity
Neurogenetics
Ion Channels, Transporters, and Neurotransmitter Reporters
Functional and Molecular Imaging
Cell Biology of Neurons, Muscles, and Glia

Additional Specifications for Video Imaging Configuration

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels
Imaging Resolution	Bi-Directional: 8 kHz Scanner: Up to 2048 x 2048 Pixels 12 kHz Scanner: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Scanner: Up to 4096 x 4096 Pixels 12 kHz Scanner: Up to 2336 x 2336 Pixels
Photoactivation	Full Field: 4-Channel
Microscope Components
Microscope Body	Z-Axis Motion
Imaging Scanners	Galvo-Resonant (8 kHz or 12 kHz)
Scan Path Wavelength Range	450 - 1100 nm
Pockels Cell	Slow (250 kHz)
Variable Attenuator	Motorized
Detection	2 Multialkali PMTs (GaAsP PMTs and Cooled PMTs Available)
Collection Optics Module	8° with Shutter
Objective Holder	Single
Objective	Olympus 20X
Sample Stage	Rigid Stand Slide Holder with XY Translation Stage
Widefield Imaging	Single-Cube Epi-Illuminator Module Four-Channel LED Source Quantalux™ sCMOS Scientific Camera
Laser	Tiberius^® fs Tunable Laser

This configuration enables video-rate, sequential two-photon imaging to study fast dynamic biological and chemical processes.

Click to Enlarge
Rigid stands provide stability for numerous sample mounting options, including the multi-camera VR theater shown here for Drosophila studies.

Click to Enlarge
Galvo-resonant scanners are available at 8 kHz and 12 kHz resonant frequencies.

Configuration B201:
Simple Z-Axis Imaging

Click for Details

Key Features

Suggested Applications

Galvo-Galvo Scanning
2-Channel Detection with High-Sensitivity GaAsP PMTs
Small Footprint with Large Throat Depth
930 nm Menlo YLMO Pulsed Laser

Neurogenetics
Neural Development and Plasticity

Additional Specifications for Simple Z-Axis Imaging Configuration

This configuration provides the simplest microscopy system for studies that require multiphoton imaging at the sample plane without unnecessary features. Please note that this configuration requires a power attenuator (not pictured).

Configuration Specifications
Highlighted System Specifications
FOV	20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed	3 fps at 512 x 512 Pixels
Scanning Resolution	Bi-Directional: Up to 2048 x 2048 Pixels Unidirectional: Up to 4096 x 4096 Pixels
Microscope Components
Microscope Body	Z-Axis Motion
Imaging Scanners	Galvo-Galvo (Ø4 mm or Ø5 mm)
Scan Path Wavelength Range	680 - 1600 nm or 450 - 1100 nm
Variable Attenuator	Manual
Detection	2 Cooled GaAsP PMTs
Collection Optics Module	8°
Objective Holder	Single
Objective	Nikon 16X
Sample Stage	Rigid Stand Slide Holder with Translation Stage
Laser	Menlo YLMO 930 nm Laser

Click to Enlarge
Menlo Systems' YLMO 930 nm laser is shown here as the primary laser source. Imaging optics may be tuned for short or long-wavelength multiphoton imaging, and the galvo-galvo scanner switched to accommodate a larger or smaller beam diameter.

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Sam Rubin
Imaging Systems General Manager

Questions?
Need a Quote?

Trans-Illumination Path Add-On for Rotating Bergamo Systems

Click to Enlarge
Trans-Illumination Add-On for Rotating Bergamo II Systems

Retrofit Options for Existing Multiphoton Systems

Upgrades to the Functionality of Existing Microscope Infrastructure and Components
Add-On Components to Increase Capabilities

Thorlabs' modular design enables our microscopes to continually evolve with experimental needs. Customers with previous model microscopes and product lines for multiphoton microscopy have the flexibility to update their infrustructure or add to their existing components as their microscopy needs change. See the expandable tables below for options available for each of our multiphoton system lines, and contact us for additional information about incorporating an upgrade or add-on into your system.

Please note that certain upgrades and add-ons require an on-site visit by one of our specialists for installation.

Bergamo II Compatible Upgrades and Add-Ons

Upgrade		Benefit		On-Site Installation Required
Scanning	SLM Holographic Photoactivation^a	Multi-Site Holographic Two-Photon Photoactivation		Yes
	Switch Scanning Functionality to Galvo-Galvo or Galvo-Resonant	Galvo-Galvo	Enable Control of Pixel Dwell Time Add Arbitrary Scanning and Photostimulation Capabilities	Yes
		Galvo-Resonant	High-Speed Imaging Capability
	12 kHz Resonant Scanner	Increase Resonant Scanning Speed		Yes
Beam Conditioning	High-Speed Pockels Cell^b	ROI Masking Using Galvo-Resonant Scanner		Yes
	Wide-Wavelength-Range Pockels Cell	Increase Operating Range to 680 - 1300 nm		Yes
Detection	10° Collection Module	Increase Emitted Signal Collection Angle Add Capability for up to 4 Channels of PMT Detection		Yes
	14° Collection Module	Increase Emitted Signal Collection Angle Increase Microscope Clearance for Electrophysiology Studies		Yes
	Mechanical Shutter for 8°, 10°, or 14° Collection Module	Protect PMTs During Photoactivation		Yes
	Polarizing Beamsplitter^c	Enable the Collection of Backscattered Multiphoton IR Signals from the Sample Plane		No
Imaging Modalities	Optics for Three-Photon Imaging	Three-Photon Excitation Capability at up to 1900 nm		Yes
	Confocal Functionality	Add Co-Registered Confocal Functionality to Primary or Secondary Path		Yes
	Convert Secondary Path Mirror to Dichroic	Enable Simultaneous Use of Epi-Fluorescence with Multiphoton Imaging		Yes
Microscope Body	Manual Condenser Focusing Module	Replace Motorized Condenser Translation with Manual Adjustment		No
	Manual Dual Objective Nosepiece	Mount Two Objectives and Manually Switch Magnification		No
	Motorized Dual Objective Nosepiece	Mount Two Objectives and Use ThorImage^®LS to Switch Magnification		No
	EMCCD/L3CCD Camera Compatibility^d	Enable the Use of an Electron-Multiplying CCD (EMCCD), or Low Light Level CCD (L3CCD) Camera		No
Support	ThorSync Hardware and Software Functionality^e	Synchronize Microscope with External Devices		No
Add-On
Beam Conditioning	Variable Beam Expander	Optimize Beam Width to the Objective Aperture Enable Deeper Penetration of Excitation Beam as Needed		Yes
	Beam Stabilizer	Compensate for Laser Angular Drift over Time Maintain Beam Position While Changing Excitation Laser Wavelength		Yes
	Variable Delay Line	Temporal Control of Two Pulsed Laser Beams		Yes
Detection	Forward PMT Detection Module	Collect Forward-Scattered Harmonic or Fluorescence Signals		Yes
	Descanned Detection	Enable Both Descanned and Non-Descanned Detection Functionality		Yes
	GaAsP PMTs	Increased Detection Sensitivity		Yes
Imaging Modalities	Sequential 2-Photon Imaging	Incorporate Tiberius^® Ti:Sapphire Femtosecond Pulsed Laser for Fast Wavelength Switching		Yes
	Transmitted Light Path for Rotating Systems	User-Removable Module Converts Rotating Microscope Between In Vivo and Slice Imaging		Yes
	Laser Scanning Dodt Contrast	Obtain Images of Sample Structure Based on Intrinsic Refractive Index Changes without Using Fluorescence		Yes
Microscope Body	Piezo Objective Mover (PFM450E)	Enable High-Resolution Z-Stack Acquisition over 450 µm Travel Range		No
	Epi-Illuminator Module with Turret for Six Filter Sets	Epi-Fluorescence Functionality with an Exchangeable Turret		No
	Scientific Camera	Add Quantalux™ sCMOS or Other Scientific CCD Cameras		No
	Translating Mounting Platform (PMP1000(/M) or PMP-2XY(/M))	Movable Breadboard Surrounding Microscope Enables Translation of Sample and/or Supplementary Equipment		No
Not Available with Galvo-Galvo Imaging Capability Requires Galvo-Resonant Scanner Available on System with 8° Detection Module Only Available Only on Rotating, Single Path System without Trinoculars May Be Added on Existing Computer Depending upon Computer Model

Bergamo I (B-Scope) Compatible Upgrades and Add-Ons

Acerra (A-Scope) Compatible Upgrades and Add-Ons

Images Taken Using Bergamo^® Systems

Selected Publications Using Thorlabs' Imaging Systems

2019

Ceriani F, Johnson SL, Sedlacek M, Hendry A, Kachar B, Marcotti W, and Mammano F. "Dynamic coupling of cochlear inner hair cell intrinsic Ca²⁺ action potentials to Ca²⁺ signaling of non-sensory cells." bioRxiv. 2019 Aug 10; 731851.

Brawek B and Garaschuk O. "Single-Cell Electroporation for Measuring In Vivo Calcium Dynamics in Microglia." Microglia Methods in Molecular Biology. 2019 Aug 8; 2034: 231-241.

Brawek B, Olmedillas del Moral M, and Garaschuk O. "In Vivo Visualization of Microglia Using Tomato Lectin." Microglia Methods in Molecular Biology. 2019 Aug 8; 2034: 165-175.

Liang Y and Garaschuk O. "Labeling Microglia with Genetically Encoded Calcium Indicators." Microglia Methods in Molecular Biology. 2019 Aug 8; 2034: 243–265.

Royzen F, Williams S, Fernandez FR, and White JA. "Balanced synaptic currents underlie low-frequency oscillations in the subiculum." Hippocampus. 2019 July 13; 23131.

Rathore APS, Mantri CK, Aman SAB, Syenina A, Ooi J, Jagaraj CJ, Goh CC, Tissera H, Wilder-Smith A, Ng LG, Gubler DJ, and St. John AL. "Dengue virus-elicited tryptase induces endothelial permeability and shock." J Clin Invest. 2019 July 2; JCI128426.

Stringer C, Pachitariu M, Steinmetz N, Carandini M, and Harris KD. "High-dimensional geometry of population responses in visual cortex." Nature. 2019 June 26; 571: 361–365.

Ziraldo G, Buratto D, Kuang Y, Xu L, Carrer A, Nardin C, Chiani F, Salvatore AM, Paludetti G, Lerner RA, Yang G, Zonta F, and Mammano F. "A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function." Front Physiol. 2019 June 11; 10: 392.

Romero S, Hight AE, Clayton KK, Resnik J, Williamson RS, Hancock KE, and Polley DB. "Cellular and widefield imaging of sound frequency organization in primary and higher-order fields of the mouse auditory cortex." bioRxiv. 2019 June 6; 663021.

Díaz-García CM, Lahmann C, Martínez-François JR, Li B, Koveal D, Nathwani N, Rahman M, Keller JP, Marvin JS, Looger LL, and Yellen G. "Quantitative in vivo imaging of neuronal glucose concentrations with a genetically encoded fluorescence lifetime sensor." J Neuro Res. 2019 May 20; 97: 946–960.

Philip V, Newton D, Oh H, Collins S, Bercik P, and Sibille E. "The Effect of Gut Microbiota on Glutamatergic/GABAergic Gene Expression in Adult Mice." Biological Psychiatry. 2019 May 15; 85: S127–S128.

Bowen Z, Winkowski DE, Seshadri S, Plenz D, and Kanold PO. "Neuronal avalanches in input and associative layers of auditory cortex." bioRxiv. 2019 Apr 29; 620781.

Burgold J, Schulz-Trieglaff EK, Voelkl K, Gutiérrez-Ángel S, Bader JM, Hosp F, Mann M, Arzberger T, Klein R, Liebscher S, and Dudanova I. "Cortical circuit alterations precede motor impairments in Huntington’s disease mice." Sci Rep. 2019 Apr 29; 9: 1–13.

Matovic S, Ichiyama A, Igarashi H, Salter EW, Wang XF, Henry M, Vernoux N, Tremblay ME, and Inoue W. "Stress-induced neuronal hypertrophy decreases the intrinsic excitability in stress habituation." bioRxiv. 2019 Mar 31; 593665.

Weissenberger Y, King AJ, and Dahmen JC. "Decoding mouse behavior to explain single-trial decisions and their relationship with neural activity." bioRxiv. 2019 Mar 5; 567479.

Ceriani F, Hendry A, Jeng JY, Johnson SL, Stephani F, Olt J, Holley MC, Mammano F, Engel J, Kros CJ, Simmons DD, and Marcotti W. "Coordinated calcium signaling in cochlear sensory and non-sensory cells refines afferent innervation of outer hair cells." The EMBO Journal. 2019 Feb 25; 38: e99839.

Lee KS, Vandemark K, Mezey D, Shultz N, and Fitzpatrick D. "Functional Synaptic Architecture of Callosal Inputs in Mouse Primary Visual Cortex." Neuron. 2019 Feb 6; 101: 421-428.e5.

2018

Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Müller JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, Cai E, Hantman AW, Wang SSH, DePiero VJ, Borghuis BG, Chapman ER, Dietrich D, DiGregorio DA, Fitzpatrick D, and Looger LL. "Stability, affinity, and chromatic variants of the glutamate sensor iGluSnFR." Nat Methods. 2018 Oct 30; 15: 936–939.

Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE,
Oertner TG, Hope BT, and Schreiter ER. "Improved methods for marking active neuron populations." Nat Commun. 2018 Oct 25; 9: 1–12.

Chen CL, Hermans L, Viswanathan MC, Fortun D, Aymanns F, Unser M, Cammarato A, Dickinson MH, and Ramdya P. "Imaging neural activity in the ventral nerve cord of behaving adult Drosophila." Nat Commun. 2018 Oct 25; 9: 1–10.

Corns LF, Johnson SL, Roberts T, Ranatunga KM, Hendry A, Ceriani F, Safieddine S, Steel KP, Forge A, Petit C, Furness DN, Kros CJ, and Marcotti W. "Mechanotransduction is required for establishing and maintaining mature inner hair cells and regulating efferent innervation." Nat Commun. 2018 Oct 1; 9: 1-15.

Saleem AB, Diamanti EM, Fournier J, Harris KD, and Carandini M. "Coherent encoding of subjective spatial position in visual cortex and hippocampus." Nature. 2018 Sept 10; 562: 124-127.

Gillet SN, Kato HK, Justen MA, Lai M, and Isaacson JS. "Fear Learning Regulates Cortical Sensory Representations by Suppressing Habituation." Front. Neural Circuits. 2018 Jan 10; 11: 112.

2017

Scholl B, Wilson DE, and Fitzpatrick D. "Local order within global disorder: synaptic architecture of visual space." Neuron. 2017 Dec 6; 95 (5): 1127-1138.

Klapoetke NC, Nern A, Peek MY, Rogers EM, Breads P, Rubin GM, Reiser MB, and Card GM. "Ultra-selective looming detection from radial motion opponency." Nature Neuroscience. 2017 Nov 09; 551: 237-241.

Kato HK, Asinof SK, and Isaacson JS. "Network-level control of frequency tuning in auditory cortex." Neuron. 2017 Jul 19; 95 (2): 412-423.

Lu R, Sun W, Liang Y, Kerlin A, Bierfeld J, Seelig JD, Wilson DE, Scholl B, Mohar B, Tanimoto M, Koyama M, Fitzpatrick D, Orger MB, and Ji N. "Video-rate volumetric functional imaging of the brain at synaptic resolution." Nature Neuroscience. 2017 Feb 27; 20: 620-628.

2016

Mongeon R, Venkatachalam V, and Yellen G. "Cytosolic NADH-NAD⁺ Redox Visualized in Brain Slices by Two-Photon Fluorescence Lifetime Biosensor Imaging." Antioxid Redox Signal. 2016 Oct 1; 25 (10): 553-563.

Pachitariu M, Stringer C, Schröder S, Dipoppa M, Rossi LF, Carandini M, and Harris KD. "Suite2p: beyond 10,000 neurons with standard two-photon microscopy." bioRxiv. 2016 Jun 30; 061507.

Dipoppa M, Ranson A, Krumin M, Pachitariu M, Carandini M, and Harris KD. "Vision and locomotion shape the interactions between neuron types in mouse visual cortex." bioRxiv. 2016 Jun 11; 058396.

Rose T, Jaepel J, Hübener M, and Bonhoeffer T. "Cell-specific restoration of stimulus preference after monocular deprivation in the visual cortex." Science. 2016 Jun 10; 352 (6291): 1319–22.

Strobl MJ, Freeman D, Patel J, Poulsen R, Wendler CC, Rivkees SA, and Coleman JE. "Opposing Effects of Maternal Hypo- and Hyperthyroidism on the Stability of Thalamocortical Synapses in the Visual Cortex of Adult Offspring." Cereb Cortex. 2016 May 26; pii: bhw096 (epub ahead of print).

Lee KS, Huang X, and Fitzpatrick D. "Topology of ON and OFF inputs in visual cortex enables an invariant columnar architecture." Nature. 2016 May 5; 533 (7601): 90-4.

Monai H, Ohkura M, Tanaka M, Oe Y, Konno A, Hirai H, Mikoshiba K, Itohara S, Nakai J, Iwai Y, and Hirase H. "Calcium imaginq reveals glial involvement in transcranial direct current stimulation-induced plasticity in mouse brain." Nat Comm. 2016 Mar 22; 7 (11100): 1-10.

Ganmor E, Krumin M, Rossi LF, Carandini M, and Simoncelli EP. "Direct Estimation of Firing Rates from Calcium Imaging Data." arXiv. 2016 Jan 4; 1601.00364 (q-bio.NC): 1-34.

2015

Roth MM, Dahmen JC, Muir DR, Imhof F, Martini FJ, and Hofer SB. "Thalamic nuclei convey diverse contextual information to layer 1 of visual cortex." Nat Neurosci. 2015 Dec 21; 19 (2): 299-307.

Barnstedt O, Keating P, Weissenberger Y, King AJ, and Dahmen JC. "Functional Microarchitecture of the Mouse Dorsal Inferior Colliculus Revealed through In Vivo Two-Photon Calcium Imaging." J Neurosci. 2015 Aug 5; 35 (31): 10927-39.

Chen SX, Kim AN, Peters AJ, and Komiyama T. "Subtype-specific plasticity of inhibitory circuits in motor cortex during motor learning." Nat Neurosci. 2015 Jun 22; 18: 1109-15.

Jia Y, Zhang S, Miao L, Wang J, Jin Z, Gu B, Duan Z, Zhao Z, Ma S, Zhang W, and Li Z. "Activation of platelet protease-activated receptor-1 induces epithelial-mesenchymal transition and chemotaxis of colon cancer cell line SW620." Oncol Rep. 2015 Jun; 33 (6): 2681-8.

Lu W, Tang Y, Zhang Z, Zhang X, Yao Y, Fu C, Wang X, and Ma G. "Inhibiting the mobilization of Ly6C^high monocytes after acute myocardial infarction enhances the efficiency of mesenchymal stromal cell transplantation and curbs myocardial remodeling." Am J Transl Res. 2015 Mar 15; 7 (3): 587-97.

Boyd AM, Kato HK, Komiyama T, and Isaacson JS. "Broadcasting of cortical activity to the olfactory bulb." Cell Rep. 2015 Feb 24; 10 (7): 1032-9.

Cossell L, Iacaruso MF, Muir DR, Houlton R, Sader EN, Ko H, Hofer SB, and Mrsic-Flogel TD. "Functional organization of excitatory synaptic strength in primary visual cortex." Nature. 2015 Feb 19; 518 (7539): 399-403.

2014

Partridge JG, Lewin AE, Yasko JR, and Vicini S. "Contrasting actions of group I metabotropic glutamate receptors in distinct mouse striatal neurones." J Physiol. 2014 Jul 1; 592 (Pt 13): 2721-33.

Peters AJ, Chen SX, Komiyama T. "Emergence of reproducible spatiotemporal activity during motor learning." Nature. 2014 Jun 12; 510 (7504): 263-7.

Ehmke T, Nitzsche TH, Knebl A, and Heisterkamp A. "Molecular orientation sensitive second harmonic microscopy by radially and azimuthally polarized light." Biomed Opt Express. 2014 Jun 12; 5 (7): 2231-46.

Liu J, Wu N, Ma L, Liu M, Liu G, Zhang Y, and Lin X. "Oleanolic acid suppresses aerobic glycolysis in cancer cells by switching pyruvate kinase type M isoforms." PLoS One. 2014 Mar 13; 9 (3): e91606.

Palmer LM, Shai AS, Reeve JE, Anderson HL, Paulsen O, and Larkum ME. "NMDA spikes enhance action potential generation during sensory input." Nat Neurosci. 2014 Feb 2; 17 (3): 383-90.

Cai F, Yu J, Qian J, Wang Y, Chen Z, Huang J, Ye Z, and He, S. "Use of tunable second-harmonic signal from KNbO₃ nanoneedles to find optimal wavelength for deep-tissue imaging." Laser & Photon Rev. 2014; 8: 865-874.

2013

Kato HK, Gillet SN, Peters AJ, Isaacson JS, and Komiyama T. "Parvalbumin-expressing interneurons linearly control olfactory bulb output." Neuron. 2013 Dec 4; 80 (5): 1218-31.

Takata N, Nagai T, Ozawa K, Oe Y, Mikoshiba K, and Hirase H. "Cerebral blood flow modulation by Basal forebrain or whisker stimulation can occur independently of large cytosolic Ca²⁺ signaling in astrocytes." PLoS One. 2013 Jun 13; 8 (6): e66525.

ThorImage^®LS Software

(Click Here for Full Web Presentation)

Features of ThorImage^®LS

Comprehensive Imaging Platform for:

Multiphoton Imaging:
- Bergamo^® II
- DIY Kits
Confocal Imaging
Hyperspectral Imaging
Multi-Modality Image Control

Seamless Integration with Experiments

Simultaneous Multi-Point Photoactivation and Imaging with Spatial Light Modulator
Fast Z Volume Acquisition with PFM450E or Third-Party Objective Scanners
Electrophysiology Signaling
Wavelength Switching with Tiberius^® Laser or Coherent Chameleon Lasers
Pockels Cell ROI Masking
Power Ramped with Depth to Minimize Damage and Maximize Signal-to-Noise

Advanced Software Functionality

Multi-Column Customizable Workspace
Image Acquisition Synced with Hardware Inputs and Timing Events
Live Image Correction and ROI Analysis
Independent Galvo-Galvo and Galvo-Resonant Scan Areas and Geometries
Tiling for High-Resolution Large-Area Imaging
Independent Primary and Secondary Z-Axis Control for Fast Deep-Tissue Scans
Automated Image Capture with Scripts
- Compatible with ImageJ Macros
Multi-User Settings Saved for Shared Workstations
Individual Colors for Detection Channels Enable Simple Visual Analysis

The full source code for ThorImage^®LS is available for owners of a Bergamo, Cerna, or confocal microscope. Click here to receive your copy.

ThorImageLS is an open-source image acquisition program that controls Thorlabs' Bergamo II, DIY Multiphoton Microscope Kits, confocal microscopes, and Cerna^® with hyperspectral imaging, as well as supplementary external hardware. From prepared-slice multiphoton Z-stacks to simultaneous in vivo photoactivation and imaging, ThorImageLS provides an integrated, modular workspace tailored to the individual needs of the scientist. Its workflow-oriented interface supports single image, Z-stacks, time series, and image streaming acquisition, visualization, and analysis. See the video at the top right for a real-time view of data acquisition and analysis with ThorImageLS.

ThorImageLS is included with a Thorlabs microscope purchase and open source, allowing full customization of software features and performance. ThorImageLS also includes Thorlabs’ customer support and regular software updates to continually meet the imaging demands of the scientific community.

For additional details, see the full web presentation.

New Functionality

Version 4.0 - October 15, 2019

Please contact ImagingTechSupport@thorlabs.com to obtain the latest ThorImageLS version compatible with your microscope. Because ThorImageLS 4.0 adds significant new features over 3.x, 2.x and 1.x versions, it may not be compatible with older microscopes. We continue to support older software versions for customers with older hardware.

New Hardware Support

Added Support for Windows^® 10 OS
Added Support for CS895MU and CS505MU Monochrome Cameras (Requires ThorCAM 3.2)
- Allows for Hot Pixel Correction
Added Support for CSN210 Motorized Dual-Objective Nosepiece
- Allows for Improved Objective Setup and Control
Added Support for Secondary Three Channel Controller
Added Support for Second LED of the DC2200 LED Driver
Added Support for New Version of Thorlabs' Tiberius^® Femtosecond Ti:Sapphire Laser
(Up to 1060 nm)
Added Support for Second Channel for GGNI (Allows for Sequential Imaging with 2 Channels)
Added Support for Controlling Up to 6 Digital Shutters (ThorShutterDig)
Added Support for Resonant-Galvo-Galvo Scan-Head (Galvo-Resonant or Gavlo-Galvo Scan Modes Only)
Added Support for Coherent^® Discovery with AOM Support (Requires Coherent^® Discovery GUI Version 1.8.3 and 3rd Party Virtual Serial Port Software)

New Features

Added Ability to Save Experiment Data in Multi-Page TIFF Format (OME TIFF)
Added Rapid Image Update for Galvo-Galvo Scanner
- Updates Image Every 16 Scan Lines During Acquisition
Added Galvo-Galvo External Trigger Sync (Minimum 1 MHz) (GGNI Not Supported)
Added Improved Galvo-Galvo and Galvo-Resonant Triggering Times
Added Ability to Read Resonant Frequency Probe
Added Configurable Trigger Output (Signal Generator) Based on Time or Other Digital Events
Added Auto Update for Histograms
Added Dedicated Bleach Shutter Control for Galvo-Galvo and GGNI
Added Stimulation Epoch Control
Added Additional Stimulation Features (Pre Idle, Post Idle) and Control Lines (Active, Cycle Output, Epoch)
Added SLM Multiple Epoch Control (Random Epoch)
Added Ability to Invert Z Control’s Plus and Minus Buttons (Supports Both Primary and Secondary Z Controllers)
Added Ability to Display X and Y Positions in Microns or Millimeters
Added BCM-PA Slider Step Size
- Allows for Setting Slide Step Size When Using Slider Plus and Minus Buttons for Power Adjustment.
Added Auto Saving of Changed Fine Alignment Values
Added Ability to Save Image Location and Zoom Level When Switching Image Modalities
Added ThorSync Changes
- Stack Panel Option
- Virtual Channel

User Interface (UI) Improvements

Renamed “Bleaching” to “Stimulation”
Added Scale Bar in Image
Added Help Menu Features
- Allows User to Check for Updates
- Allows User to View Log File for Trouble-Shooting
Added Shortcuts to Hardware Settings and Application Settings in Hardware Connections Window and Edit Under Settings Menu
Changed Capture Preview of Image to Show Averaged if Cumulative Mode is Used
Added Control Digital Switches within Script
Updated Digital Switch Configuration to be Saved in Experiment Settings and Viewable in Experiment Settings Browser
Updated Extend Filing Numbering Index Out to 6 Digits

Fixed Bugs

Fixed - Fast Switching was not Disabled When Hiding Control for Tiberius fs Ti:Sapphire Laser

Brochures and Mind Map

The buttons below link to PDFs of printable materials for Bergamo^® II microscopes.

Laser Scanning
Scan Path Wavelength Range		450 - 1100 nm, 680 - 1600 nm, or 900 - 1900 nm
Scan Paths		Resonant-Galvo-Galvo Scanner, Galvo-Resonant Scanners, Galvo-Galvo Scanners, or Spatial Light Modulator; Single or Dual Scan Paths
Scan Speed	8 kHz Resonant-Galvo-Galvo or Galvo-Resonant	2 fps at 4096 x 4096 Pixels 30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels
	12 kHz Resonant-Galvo-Galvo or Galvo-Resonant	4.4 fps at 2048 x 2048 Pixels 45 fps at 512 x 512 Pixels 600 fps at 512 x 32 Pixels
	Galvo-Galvo	3 fps at 512 x 512 Pixels 48 fps at 512 x 32 Pixels 70 fps at 32 x 32 Pixels Pixel Dwell Time: 0.4 to 20 µs
Galvo-Galvo Scan Modes		Imaging: Line, Polyline, Square, or Rectangle Non-Imaging: Circle, Ellipse, Polygon, or Point
Field of View		20 mm Diagonal Square (Max) at the Intermediate Image Plane [12 mm Diagonal Square (Max) for 12 kHz Scanner]
Scan Zoom		1X to 16X (Continuously Variable)
Scan Resolution		Up to 2048 x 2048 Pixels (Bi-Directional) [Up to 1168 x 1168 Pixels for 12 kHz Scanners] Up to 4096 x 4096 Pixels (Unidirectional) [Up to 2336 x 2336 Pixels for 12 kHz Scanners]
Compatible Objective Threadings		M34 x 1.0, M32 x 0.75, M25 x 0.75, and RMS

Multiphoton Signal Detection
Epi-Detection	Up to Four Ultrasensitive GaAsP PMTs, Cooled or Non-Cooled
Forward-Direction Detection	Two Ultrasensitive GaAsP PMTs
Maximum of Four PMTs Controlled by the Software at a Given Time
Collection Optics	8°, 10°, or 14° Collection Angle (Angles Quoted When Using an Objective with a 20 mm Entrance Pupil) Easy-to-Exchange Emission Filters and Dichroic Mirrors

Confocal Imaging
Motorized Pinhole Wheel with 16 Round Pinholes from Ø25 µm to Ø2 mm Two to Four Laser Lines (488 nm Standard; Other Options Range from 405 nm to 660 nm) Standard Multialkali or High-Sensitivity GaAsP PMTs Easy-to-Exchange Emission Filters and Dichroic Mirrors

Widefield Viewing
Manual or Motorized Switching Between Scanning and Widefield Modes Illumination Provided via LED or Liquid Light Guide C-Mount Threads for Scientific Cameras

Transmitted Light Imaging
Differential Interference Contrast (DIC) or Dodt Gradient Contrast Widefield or Laser Scanned Illumination Provided by Visible and/or NIR LEDs Compatible with Air or Oil Immersion Condensers

Three-Photon Imaging
Scan Optics for 900 - 1900 nm Range Achieve Reduced Background Scatter for Greater Sensitivity in Deep Tissue Imaging

Volume Imaging Using Bessel Beams
3D Volumetric Functional Imaging at Video Frame Rates Enhanced Temporal Resolution for Studying Internal Systems at Cellular Lateral Resolution In Vivo

Translation
Microscope Body Rotation (Rotating Bodies Only)	-5° to +95°, -50° to +50°, or -45° to +45° Around Objective Focus 0.1° Encoder Resolution
Coarse Elevator Base Z (Rotating Bodies Only)	5" (127 mm) Total Travel; 1 µm Encoder Resolution
Fine Microscope Body X and Y	2" (50.8 mm) Total Travel; 0.5 µm Encoder Resolution
Fine Microscope Arm Z	1" (25.4 mm) Total Travel; 0.1 µm Encoder Resolution
Fine Objective Z (Piezo Objective Scanner)	Open Loop: 600 µm ± 10% Travel Range; 1 nm Resolution Closed Loop: 450 µm Travel Range; 3 nm Resolution

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Posted Comments:

jfpena (posted 2016-12-19 18:15:55.003)

I am looking for a cheap way to do confocal imaging in vivo. Is this Bergamo II Series Multiphoton Microscope my best option? Can you send me a quote?

tfrisch (posted 2016-12-22 11:44:31.0)

Hello, thank you for contacting Thorlabs. A member of our Imaging Team will reach out to you directly to discuss this system and your application.

birech (posted 2016-11-17 06:33:49.463)

I asked for a price quote for this product, Bergamo II Series Multiphoton Microscopes three days ago. I am working at the University of Nairobi in Kenya and would wish to order one. Regards, Birech

tfrisch (posted 2016-11-17 06:56:23.0)

Hello, thank you for contacting Thorlabs. I have forwarded this request to our Imaging Sales Team. I apologize for the delay.

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Bergamo® II Series Multiphoton Microscopes

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Imaging Systems Demo Rooms

Options at a Glance

Innovation through Collaboration

Five-Axis Movement of Rotating Bergamo Microscopes

Features

Thorlabs recognizes that each imaging application has unique requirements.If you have any feedback, questions, or need a quotation, please use our multiphoton microscopy contact form or call (703) 651-1700.

Bergamo® II Modules

Galvo-Resonant Scanners, Galvo-Galvo Scanners, and Spatial Light Modulators

Fast Switching Using a Tunable Femtosecond Laser

Volume Imaging Technique Using Bessel Beams

Periscopes

Super Broadband Scan Optics

Large-Angle Signal Collection Optics

Easy-to-Reach Emission Filters and Dichroic Holders

Detectors in Epi and Transmitted Directions

Multi-Axis Controller with Touchscreen

Objectives

Rigid Stand Sample Holders

Scientific Cameras

User-Installable Dodt Contrast and DIC Imaging Modules

Thorlabs recognizes that each imaging application has unique requirements.If you have any feedback, questions, or need a quotation, please use our multiphoton microscopy contact form or call (703) 651-1700.

Structural Neurobiology

Neurological Disorders

Neural Development and Plasticity

Neurogenetics

Functional and Molecular Imaging

Synapses and Circuits

Ion Channels, Transporters, and Neurotransmitter Reporters

Cell Biology of Neurons, Muscles, and Glia

Drug Discovery

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Application Summary

Publications

Example Configurations

Additional Specifications for Multi-Target Photoactivation Configuration

Additional Specifications for 2P / 3P Imaging Configuration

Additional Specifications for Random Access Scanning Configuration

Additional Specifications for In Vivo 2P Imaging Configuration

Additional Specifications for Dual-Path Random Access Scanning Configuration

Additional Specifications for Dual-Path Confocal Configuration

Additional Specifications for Simple XYZ Configuration

Additional Specifications for Video Imaging Configuration

Additional Specifications for Simple Z-Axis Imaging Configuration

Thorlabs recognizes that each imaging application has unique requirements.If you have any feedback, questions, or need a quotation, please use our multiphoton microscopy contact form or call (703) 651-1700.

Retrofit Options for Existing Multiphoton Systems

Images Taken Using Bergamo® Systems

Selected Publications Using Thorlabs' Imaging Systems

2019

2018

2017

2016

2015

2014

2013

ThorImage®LS Software

(Click Here for Full Web Presentation)

New Functionality

Brochures and Mind Map

Thorlabs recognizes that each imaging application has unique requirements.If you have any feedback, questions, or need a quotation, please use our multiphoton microscopy contact form or call (703) 651-1700.

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Bergamo^® II Modules

Fast Switching Using a
Tunable Femtosecond Laser

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.

Images Taken Using Bergamo^® Systems

ThorImage^®LS Software

Thorlabs recognizes that each imaging application has unique requirements.
If you have any feedback, questions, or need a quotation, please use our
multiphoton microscopy contact form or call (703) 651-1700.