How to read the results of fluorescent microscopy using tunel to detect apoptosis
1. Pay attention to three points in the observation of the results, the epi-projection method, the resolution projection and the filter treatment. Specifically:
The illumination method is usually episcopic, that is, the light source is projected on the sample through the objective lens;
The light source is ultraviolet light, the wavelength is shorter, and the resolution is higher than that of ordinary microscopes;
There are two special filters, the one in front of the light source is used to filter out visible light, and the one between the eyepiece and objective lens is used to filter out ultraviolet rays to protect human eyes.
Second, the fluorescent microscope used to measure apoptosis by tunel is also a kind of optical microscope, the main difference is that the excitation wavelengths of the two are different. This determines the difference between the fluorescence microscope and the ordinary optical microscope in terms of structure and usage. Fluorescence microscopy is an essential tool in immunofluorescent cytochemistry. It is composed of main components such as light source, filter plate system and optical system. It is to use a certain wavelength of light to excite the specimen to emit fluorescence, and to observe the fluorescence image of the specimen by amplifying the objective lens and eyepiece system.
3. Fluorescence suppressed filter plate for measuring apoptosis in tunel The function of the suppressed filter plate is to completely block the passage of excitation light and provide fluorescence in the corresponding wavelength range. Corresponding to the excitation filter plate, the following three types of pressed filter plates are commonly used:
Ultraviolet light pressing filter plate: It can pass visible light and block ultraviolet light from passing through. Can be combined with UG-1 or UG-5. Commonly used GG-3K430 or GG-6K460.
Violet blue light suppression filter plate: can pass light with a wavelength above 510nm (green to red), and can be combined with BG-12. Usually OG-4K510 or OG-1K530 is used.
Ultraviolet light pressing filter plate: can pass light with a wavelength above 460nm (blue to red), can be combined with BG-3, commonly used OG-11K470AK 490, K510.
