Open the microscope and calibrate the optical path. Focus on the relevant area of the specimen. Check the connection between the camera and the computer, direct the light to the camera, focus, and save the image.
Operation method
Program 16.6 Digital Photography of Microscopes
Principle
Open the microscope and calibrate the optical path. Focus on the relevant area of the specimen. Check the connection between the camera and the computer, direct the light to the camera, focus, and save the image.
Materials and Instruments
Cultures to be examined Move 1. Prepare cultures for photography (ensure that cultures are free of debris as in the case of replacing primary cultures prior to photography). For more product details, please visit Aladdin Scientific website.
Inverted microscope, lens paper, image recording paper, micrometer.
(a) Bottle culture
i) Take the culture bottle to the ultra-clean table, loosen the cap and cool the culture.
ii)When the temperature of the culture is room temperature, close the cap tightly. Deformation caused by cooling can be avoided.
iii)Turn the culture to rinse the inside surface of the top of the culture bottle to remove any condensation that may have formed.
iv)Stand the culture bottle upright to allow the culture solution to flow down.
(b) Petri dish or culture plate cultures
i)In an ultra-clean bench, cool the culture.
ii)Replace the lid of the petri dish or culture plate (make sure the sample marker is on the bottom of the petri dish or culture plate).
2. Select the field of view and magnification (4× is best for clonal or monolayer cell arrangements; 10× for typical pictures; 20× for pictures of cellular details), avoiding imperfections or markings on the culture flask (always mark the side of the flask or petri dish, not the top or bottom). (The inverted microscope is equipped with the following accessories: 10×, 20×, or 40× aberration objectives; a condenser with a phase-aberration ring for 10×, 20×, or 40× objectives; a trinocular head or other interface for CCD cameras; and neutral density filters, 2× and 4×.)
3. Checking the focal length
(a) Adjust the focal length of both eyepieces to fit your eyes using the lines in the viewfinder or objective lens;
(b) Focus the microscope.
4. Turn on the monitor and examine the density and contrast of the image on the screen. Adjust the intensity of the microscope light using a rheostat for black-and-white photographs and a neutral density filter for color photographs (if the color temperature is incorrect, the photographs can be processed electronically at a later stage).
5. Refocus the microscope if required.
6. store the photographs at a resolution suitable for the end use and permitted by the storage device (e.g., 940 × 740 requires 2 Mb of storage space, which is the right quality if 5 × 7 inches, i.e., 12 cm × 18 cm, images are developed).
7. Turn down the power supply brightness to prevent the culture from overheating. (An infrared filter can be used to minimize overheating.)
8. Repeat steps 3-6 with a micrometer at the same magnification to obtain the magnification. If the image is processed in the same way, it can be used as a scale.
9. Place the culture back into the incubator.
10. Complete the documentation of what was captured in the image according to the saved file name, otherwise the image will be illegible.
11. Images can be stored, viewed, edited, or formatted using the Adobe PhotoShop program, or they can be printed using a high-quality inkjet or color laser printer and Kodak ColorEase.