Hi, I am taking pictures of Immunohistochemistry slides using microscope attached camera. At high magnification x400 i am getting yellow tinted pictures can any body expertise in microscopy or photography troubleshoot the problem. Thank you.
Mir Khurshid Iqbal
Experimental cancer biochemistry.
If you want consistency with a tungsten lamp use it at the same power setting as much as you can each time. The easiest setting for that is full power. The temperature of the filament will create the colour you are imaging with. If you have a decent microsopcy camera on your microscope & the software it came with it likely has an option to adjust white balance. These are also usually calibrated to work with a tungsten filament at 3200K.
To test this is set up correctly insert an empty slide in to the microscope at the same focus as a sample (stick real sample in, set it up then swap to the empty one) and check your images are evenly grey all over. This is the perfect test to ensure you have consistently beautiful images for IHC. the testing approach will also work with most photography cameras as you can check first! As Sönke said there are white balance options on these as well.
Dear Khurshid,
That sounds to me that your white balace is off. You should check if the microscope is equiped with a blue filter in front of the the lamp and the condensor. Also set your camera (assuming that it is a consumer type camera - non microscopic one) to a white balance to tungsten light.
That should give you better images.
Best Soenke
Also check that you microscope do not have additional filters (in general they have yellow filters)
If you want consistency with a tungsten lamp use it at the same power setting as much as you can each time. The easiest setting for that is full power. The temperature of the filament will create the colour you are imaging with. If you have a decent microsopcy camera on your microscope & the software it came with it likely has an option to adjust white balance. These are also usually calibrated to work with a tungsten filament at 3200K.
To test this is set up correctly insert an empty slide in to the microscope at the same focus as a sample (stick real sample in, set it up then swap to the empty one) and check your images are evenly grey all over. This is the perfect test to ensure you have consistently beautiful images for IHC. the testing approach will also work with most photography cameras as you can check first! As Sönke said there are white balance options on these as well.
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