Vitamin C crystals in polarized light taken with the Moticam ProS5 Lite camera 100X© Robert Berdan
Moticam ProS5 Overview
The Moticam ProS5 Lite dedicated microscope camera offers 5MP (2448 x 2048 px) and can be attached to a microscope via a C-mount. A C-mount is a type of lens mount commonly found on 16 mm movie cameras, machine vision cameras, and microscope phototubes. This camera can also use a special eyepiece for insertion into an ocular tube of a compound or stereoscope microscope. C-mount eyepieces vary in magnification from 0.35X to 1.0X and it’s important to choose the correct eyepiece depending upon the camera’s sensor. The Moticam ProS5 Lite uses a 2/3” sensor with 3.45 x 3.45 μm pixels. If you plan to purchase an eyepiece or a C-mount adapter for the Moticam, contact a Motic sales representative so they can recommend the correct adapter or eyepiece. I spent a week testing the Moticam ProS5 Lite camera with a Motic BA310 polarizing microscope.
The Moticam ProS5 Lite uses a global shutter. Global shutters allow all of the pixels to accumulate a charge with the exposure starting and ending at the same time. At the end of the exposure time, the charge is read out simultaneously. Therefore, the image shows no motion blur with moving subjects (J. Chouinard, 2018). The Moticam ProS5 Lite camera uses a 2/3 inch sCMOS (scientific Complementary metal-xxide-xemiconductor) chip. Live display mode is available via a USB connection to a computer. The camera supports both USB 2 and 3.1. The camera offers exposures between 14 microseconds to 2 seconds. To learn more about Moticam cameras and their pricing, check Motic camera specifications and pricing page on the web site. The Moticam ProS5 Lite camera costs about $1000.
Installing The Moticam Software: A Step-by-Step Guide
Step 1. Insert the USB stick with Moticam software into the USB port of your computer.
Step 2. View the files on the USB stick. On Windows right-click on the Motic icon and select open and you will see Linux, Mac, and Windows folders. Open the folder that has software for your operating system. I selected the Windows x64 folder for installing on a 64-bit operating system. Then double-click on the setup.exe file to initiate the installation and follow the on-screen instructions (on some computers you may also have to install the Twain driver).
Step 3. Put a microscope slide and specimen on the microscope stage with the microscope light turned on and focused on the specimen. If using a trinocular head attached to the camera pull out the trinocular head slider or rotate the head (with some microscopes) to illuminate the camera.
Step 4. Plug the Moticam camera USB cord into the computer and start the Moticam software. During Installation the installer software puts an icon on your computer desktop, if you don’t see it look in the programs folder for the moticam.exe file to start the program. The camera has a small blue LED (light-emitting diode) that indicates it has power provided by the computer.
Vitamin C crystals in polarized light 100X© Robert Berdan
Step 5: Below is the first screen image you will see when you start the Moticam software.
Step 6: To view the camera image, select File > Capture in the software as shown below or use the keys Ctrl + M.
Step 7: Once you select file capture you should see a screen like that shown below with your specimen. If the screen appears dark or white the image may be under or overexposed or you may have the trinocular head slider in the incorrect position. It’s likely that the exposure and gain will need to be adjusted. Select the first button on the right side of the screen and adjust the exposure and gain sliders to achieve a correctly exposed image. The first button on the right side of the screen permits control of the camera exposure and sensor gain by using the sliding buttons.
Step 8: Select the next button down on the right side of the window to adjust the individual RGB colors. Look for a grey area in the specimen and try to make it neutral in color or match the colors to what you see in the microscope. There are no auto white balance or auto color options. Having a color-calibrated computer monitor is helpful but not essential. Once you determine the optimum settings for different types of microscope lighting I recommend writing them down in a notebook for future reference. The 2nd button down on the right panel takes to Colour adjustment controls.
Step 9: In the below screengrab, note the 4th button down on the right side with a camera icon. When you select this button the controls permit you to capture a single image or record video (up to 2448 x 2048 37.5 fps). Video is saved as a moviefilename.avi on a PC. The 5th button down on the right side allows you to calibrate the software to make morphometric measurements and add scale bars to the images. See the online manual for instructions on how to do this. You need to calibrate each objective. The right-side camera icon button takes to image and video image capture screen.
Step 10: The ability to zoom into the computer screen will be helpful to achieve critical focus. To do this choose the ROI (Region Of Interest) preview radio button in the top menu and drag a rectangular selection with the mouse (click and drag) over a small part of the image and then chose a zoom setting. Then focus carefully with the microscope fine focus until the image is sharp. After this turn off the ROI button and delete the square box (select the box and hit delete or click on the picture outside the box) and then capture an image. If you don't remove the ROI red box it will appear on the final picture.
Below are the settings I used to take photomicrographs of crystals in polarized light using the Moticam ProS5 Lite and the Motic BA310 polarizing microscope:
|Microscope Illumination||Full Power|
The settings above will vary depending on the microscope, camera model, filters, the sample, and the type of illumination you are using.
Step 11: After closing the “top” overlying window by selecting the X at the top of the right panel you should see the initial software screen shown below with one or more image thumbnails at the bottom of the screen. Select a thumbnail and it will appear large in the main window. Before you save each image you need to select the size of the saved image and a folder where you want to save the image file. You can save the file as a .jpg or .tif file. The below screengrab shows the initial software window after taking a single picture with the Moticam.
Step 12: After closing the window used to capture images, you will see the images you took as thumbnails at the bottom of the screen. When you select an image thumbnail you will see a large photo appear above it. You then need to choose an image size before saving - if you don't do this the image will default to 1072 x 764 pixels. At the top of the screen Select > Image > Image size > choose an image size to save the file. After saving each image close the large image window (not the program window) by clicking on X at the top right of each image pane. Then navigate to the folder where you saved the images to work with the files or open them in another image editing program for further processing. When you save the image it also saves a text file with it (filename.tif.mwi). MWI text files can be opened in Windows notepad and contain information additional information about the image.
Moticam software can save image files at a maximum size of 2448 x 2048 pixels at 72 dpi resulting in a 14.3 MB file. This size image will allow you to make a print - 6.7 x 8 inches at 300 dpi. There are other features in the software which can be used to measure objects, paint, add text, or create albums. To learn more about these features read the online manual under help. The camera includes a microscope slide with calibration objects (dot, bar, and ruler) that can be used with the software to add scale bars on images taken with different magnification objectives, determine object sizes, and areas for morphometric analysis.
Pinewood section viewed by polarized light and full-wave compensator 400X© Robert Berdan
Calanoid copepod- side view by Polarized light microscopy. Note the single red eye and birefringent muscles. 100X.© Robert Berdan
Epsom crystals (magnesium sulfate) polarized Light microscopy 100X.© Robert Berdan
Vitamin C crystal Polarized light microscopy 100X.© Robert Berdan
Cladoceran Ceriodaphnia sp with eggs - polarized light microscopy using a full 550 nm compensator 100X.© Robert Berdan
To make larger images with this camera you can photograph overlapping regions and stitch them together with software. Motic offers software called EasyStitchPro that automatically stitches images, but there are many image stitching programs to choose from. I often use Photoshop for image stitching, focus stacking, and image enhancement.
Motic offers a variety of digital cameras for their microscopes starting around $400. Dedicated cameras have some advantages over DSLRs (Digital Single Lens Reflex cameras) including their compact size, lack of vibration during exposure, and the ability to process the image by manipulating exposure, color, and gain before taking the photo. The software also offers the ability to carry out basic image analysis of the specimen and quantification of the image data.
A panoramic image created by stitching 24 images together of Vitamin C crystals photographed with polarized light microscopy with a 10X objective© Robert Berdan
The bone section shown in polarized light and a full-wave compensator 100X.© Robert Berdan
A & B ) Vitamin C crystals polarized light C) Wool fibers polarized light D) Vitamin C crystal Polarized light E) Potato starch grains polarized light F) Potato starch grains polarized light and full-wave 550 nm compensator. All taken at 100X.© Robert Berdan
References & Links:
1. J. Piper and R. Pelc (2020) Consumer Versus Dedicated Digital Cameras in Photomicrography. Chapt 13. In book: Neurohistology and Imaging Techniques (Neuromethods, vol. 153) Publisher: Springer (Humana Press)
2. R. Berdan and B. Berdan (2020) Cell Phone Cameras, Dedicated Digital Cameras, and Digital Single Lens Reflex Cameras for Photomicrography.
3. Motic EasyStitch Pro Software – PDF https://www.canadiannaturephotographer.com/MoticPolscope/EasyStitch%20Pro%20flyer%20pricing-compressed.pdf
4. Moticam ProS5 Lite Camera specifications and pricing on Motic North America Web site
5.J. Chouinard (2018) Global vs Rolling shutters.