Pre examination and adjustment of fluorescence microscopy:
(1) Before each fluorescence observation, it is necessary to routinely check the filament alignment, optical path focusing, aperture aperture aperture, and field of view aperture settings of the fluorescence device.
(2) Whether the required fluorescence excitation/emission filter components are installed in the converter, whether the fluorescence microscope objective is configured properly, and whether the oil stains and dust in front of the objective lens are removed.
(3) Similarly, when observing the difference in transmitted light, it is necessary to check the conjugation of the focusing lens to the center and the contrast ring to the objective lens.
(4) Check if there is any liquid or dust hanging on the sample carrier (glass slide, cover glass, and other vessels), and if the thickness is within the working distance range calibrated by the objective lens. Sliced samples should not be too thick, approximately ≤ 10 μ m is recommended.
(5) Due to the presence of ultraviolet rays in the lighting source, a brown sunshade should be placed above the loading platform to prevent UV damage to the retina.
(6) Unstable voltage can reduce the service life of high-voltage mercury lamps, and a voltage regulator should be added to the light source power supply.
(7) To extend the lifespan of the mercury lamp, it can only be turned off 15 minutes after being turned on; Once the fluorescent power supply of the mercury lamp is turned off, it needs to wait for at least 10 minutes to cool the mercury vapor back to its original state when restarted, otherwise it will affect the lifespan of the lamp.
Observation of fluorescence microscope images:
(1) After turning on the fluorescent lamp source for about 5-10 minutes, the excitation intensity tends to stabilize, and the sample is loaded for observation; To prevent excessive excitation of light during focusing and searching for images from causing fluorescence quenching of the sample, the aperture of the fluorescence microscope is first reduced or an ND filter is added to adjust the excitation to a moderate intensity, and the sample stage is moved regularly. After determining the mirror image, it is adjusted to the fluorescence state for recording.
(2) Adjustments with poor image quality. The necessary adjustment measures that can be taken, excluding sample preparation factors, are:
① Exclude shading or limiting devices in the imaging optical path, such as DIC accessories, ND filters, etc.
② Readjust the focusing and aperture aperture aperture of the fluorescence microscope's collector.
③ Carefully adjust the coverage correction ring of the fluorescence microscope objective.
