Two-Camera Mounts for Microscopes

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Figure 1.1  This image shows a live, simultaneous overlay of fluorescence and near-infrared Dodt contrast images of a 50 µm brain section from a CX3CR1-GFP mouse. More details can be found on the Applications tab. Sample courtesy of Dr. Andrew Chojnacki, University of Calgary.

Features

• Microscope Adapters Allow Two Scientific Cameras to Image a Single Optical Input (See the Applications Tab for Examples)
• Ideal for Calcium Ratio Imaging and Electrophysiology
• Accepts 25 mm x 36 mm Dichroic Filters or Beamsplitters
• Accepts Standard Ø25 mm or Ø1" Filters on Outputs
• Fine Pitch Rotation and XY Adjustment for Image Registration
• Coarse Focus Adjustment for Parfocalization
• Adapts to Most Cerna, Olympus, and Nikon Upright Microscopes Using Thorlabs' Standard SM1 Interface (Microscope Camera Port Adapters Sold Separately)
• Supported by ThorCam™ Image Overlay Plug-In for Our Scientific Cameras

These two-camera mounts are designed to attach two Thorlabs scientific cameras to an upright Cerna, Olympus, or Nikon microscope, allowing simultaneous imaging of a single optical output. Typical applications include multispectral imaging using a dichroic beamsplitter; for more details please see the Applications tab. A rotation mount allows for 360° of rotational adjustment (±8° fine adjustment) for the reflected camera while a translation mount gives 4 mm linear XY adjustment of the transmitted camera. Both camera mounts have coarse focus adjustment by manually translating the cameras, allowing for parfocalization of both images. The 2CM1 and 2CM2 mounts have up to 15 mm and 11 mm of adjustment, respectively, using the cage rods, although this adjustment range may be limited by the geometry of the camera's front face.

The 2CM1 mount is designed for cameras with 60 mm cage system taps on the front, such as our cooled sCMOS and CMOS cameras, while the 2CM2 mount is identical except for the inclusion of two LCP4S cage size adapters for compatibility with 30 mm cage system taps, such as those found on our compact scientific cameras with sCMOS and CMOS sensors.

Each mount includes a DFM1 fluorescence filter cube, which is designed to hold a fluorescence filter set (dichroic mirror, excitation filter, and emission filter) as well as 25 mm x 36 mm plate beamsplitters or other optics (up to 1.1 mm thick). The filter cube consists of a base and top lid with an insert to hold filter set components and has a kinematic design for easy swapping between mounted filter sets without requiring realignment. Additional DFM1T1 top lids for mounting different filter sets are sold separately below. See the Applications tab for example uses.

Microscope Compatibility
The input port of each two-camera mount has external SM1 (1.035"-40) threading that can mate with our SM1-threaded microscope camera port adapters for installation on many commercial upright microscopes. The 2CM1 and 2CM2 mounts place our scientific cameras' sensors at a distance of 4.05" to 4.17" (102.9 mm to 106.0 mm) from the base of the mount, which may be outside the parfocal distance of some microscopes, including inverted microscopes from Nikon and Olympus. For these microscopes, our two-camera mounts can be used if parfocality with the eyepieces is not needed.

These mounts will be parfocal with Thorlabs' Cerna^® Series Microscopes with our previous-generation inverted-image trinoculars, as well as upright microscopes from Nikon and Olympus. For Cerna Series Microscopes with inverted-image trinoculars and upright Nikon Eclipse microscopes, these mounts can be attached to the trinoc camera port using our SM1A58 camera port adapter. For Olympus BX and IX microscopes, Thorlabs offers the SM1A51 camera port adapter with SM1 threading. These mounts are not compatible with our upright-image trinoculars.

Turnkey prealigned systems with cameras and three-way camera mounts are also available as custom items. Please contact Tech Support for more details.

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Figure 1.2  Screenshot of ThorCam Overlay Plug-in

Compatible with Thorlabs' Scientific Cameras

• High Quantum Efficiency
• Low Read Noise
• Polarization Camera Available
• Triggered and Bulb Exposure Modes
• ThorCam GUI included
• Third-Party Software Support for LabVIEW^®, MATLAB^®, µManager, and Metamorph^®

These two-camera mounts are ideal for use with Thorlabs' scientific cameras, which have four 4-40 tapped holes on the front camera face for mounting to the two-camera mounts.

Thorlabs' scientific cameras are based on high quantum efficiency, low-noise imagers, which make them ideal for multispectral imaging, fluorescence microscopy, and other high-performance imaging techniques. Both non-cooled and cooled versions are available. For more details about our camera families, please click on the links in Table 1.3.

The ThorCam user interface provided with our scientific cameras includes a plug-in to allow for two live camera images to be overlaid into a real-time 2-channel composite, eliminating the need for frequent updates of a static overlay image. This live imaging method is ideal for applications such as calcium ratio imaging and electrophysiology. As seen in Figure 1.2, the plug-in controls image threshold and opacity (Alpha) settings, and can apply false color to one of the two monochrome channels. Please refer to the Applications tab for more examples.

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Figure 2.1  Example Setup for Simultaneous NIR/DIC and Fluorescence Imaging

Live Dual-Channel Imaging

Many life science imaging experiments require a cell sample to be tested and imaged under varying experimental conditions over a significant period of time. One common technique to monitor complex cell dynamics in these experiments uses fluorophores to identify relevant cells within a sample, while simultaneously using NIR or differential interference contrast (DIC) microscopy to probe individual cells. Registering the two microscopy images to monitor changing conditions can be a difficult and frustrating task.

Live overlay imaging allows both images comprising the composite to be updated in real-time versus other methods that use a static image with a real-time overlay. Overlays with static images require frequent updates of the static image due to drift in the system or sample, or due to repositioning of the sample. Live overlay imaging removes that dependency by providing live streaming in both channels.

Using the ThorCam overlay plug-in with the two-way camera microscope mount, users can generate real-time two-channel composite images with live streaming updates from both camera channels, eliminating the need for frequent updates of a static overlay image. This live imaging method is ideal for applications such as calcium ratio imaging and electrophysiology.

Example Images

Simultaneous Fluorescence and DIC Imaging

Figures 2.2 and 2.3 show mouse kidney cells imaged using a dichroic filter to separate the fluorescence and DIC signals into different cameras. These images are then combined into a two-channel composite live image with false color fluorescence by the ThorCam overlay plug-in (see Figure 2.4).

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Figure 2.2  Fluorescence Image

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Figure 2.3  DIC Image

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Figure 2.4  2-Channel Composite Live Image

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Figure 2.5  In the image above, the pipette is visible in the DIC image as two lines near the center of the frame.

Microaspiration Using a Micropipette

Figure 2.5 shows a live, simultaneous overlay of fluorescence and DIC images. The experiment consists of a microaspiration technique using a micropipette to isolate a single neuron that expresses GFP. This neuron can then be used for PCR. This image was taken with our previous-generation 1.4 megapixel cameras and a two-camera mount and shows the live overlay of fluorescence and DIC from the ThorCam plug-in. Image courtesy of Ain Chung, in collaboration with Dr. Andre Fenton at NYU and Dr. Juan Marcos Alarcon at The Robert F. Furchgott Center for Neural and Behavioral Science, Department of Pathology, SUNY Downstate Medical Center.

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Figure 2.6  This image shows a live, simultaneous overlay of fluorescence and near-infrared Dodt contrast images of a 50 µm brain section from a CX3CR1-GFP mouse. More details can be found on the Applications tab. Sample courtesy of Dr. Andrew Chojnacki, University of Calgary.

Simultaneous NIR Dodt Contrast and Epi Fluorescence imaging

Figure 2.6 shows a live, simultaneous overlay of fluorescence and near-infrared Dodt contrast images of a 50 µm brain section from a CX3CR1-GFP mouse, which has been immunostained for PECAM-1 with Alexa-687 to highlight vasculature. The Dodt contrast uses a quarter annulus and a diffuser to create a gradient of light across the sample that can reveal the structure of thick samples. The image was taken with our scientific cameras and a two-camera mount. Sample courtesy of Dr. Andrew Chojnacki, Department of Physiology and Pharmacology, Live Cell Imaging Facility, Snyder Institute for Chronic Diseases, University of Calgary.

Other Configurations

The 2CM1 and 2CM2 Two-Camera Mounts can also be utilized with other optics sharing the same 25 mm x 36 mm x 1 mm size of a standard dichroic mirror. Figure 2.7 suggests a two-camera imaging system designed for dual wavelength fluorescence imaging with a dichroic mirror. In Figure 2.8, a 50:50 plate beamsplitter can be used to split the signal evenly; this is useful, for example, for imaging the sample using two different cameras simultaneously.

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Figure 2.8  Example Setup for Dual Wavelength Fluorescence Imaging Using a 50:50 Plate Beamsplitter

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Figure 2.7  Example Setup for Dual Wavelength Fluorescence Imaging Using a Dichroic Mirror