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Fluorescence microscope use method and operation procedure

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Fluorescence microscope use method and operation procedure

Date of release:2019-08-07 Author: Click:

1. Cover the light with curtains, turn off the light in the room, remove the dust cover of the microscope, and ensure that the microscope lamp room is well ventilated and uncovered. Install the mercury lamp light box and turn the lever on the light box collar to connect the light box to the arm. Coal and rock analysis system

2. Insert the power plug of the mercury lamp light box into the socket behind the fluorescent power box, and then plug the plug of the fluorescent power box into the 220V external power supply.

3. Turn on the power switch, the voltmeter shows the power supply voltage. If the power supply voltage fluctuation is not more than ±5% of the rated voltage value, press the start switch to ignite the mercury lamp. For example, because the weather is too cold or the voltage is unstable, etc. Start the unignited mercury lamp and you can move it a few more times. When the arc of the ultra-high pressure mercury lamp reaches a steady state and reaches the maximum luminous efficiency, it can start working.

4. The adjustment of the bulb:

(1) An optional specimen is placed on the stage;

(2) Rotating the condenser knob on the mirror arm to move the condenser out of the optical path, rotating the color filter set to convert the hand wheel, and rotating the violet (V) or blue (B) or green (G) excitation filter set into the optical path, and Transfer the 10X fluorescent objective lens into the optical path;

(3) Adjust the coarse and fine adjustment hand wheel to make the specimen image focus clear;

(4) Push the focusing lens on the right side of the vertical illuminator back and forth to make the field light bar image clear, turn the field light bar to close the field light bar, adjust the field light bar to adjust the middle view screw to make the field of view The light bar is centered, and then the field diaphragm is opened to the maximum;

(5) Rotating the condenser knob on the mirror arm to move the condenser into the optical path, and adjusting the condenser lens lever on the light box before and after, so that the arc of the mercury lamp is clearly visible in the field of view;

(6) Adjust the bulb level adjusting screw and the vertical adjusting screw on the light box to center the arc of the mercury lamp;

(7) Adjust the mirror horizontal adjustment screw and the vertical adjustment screw to separate the reflection image of the light source from the arc of the mercury lamp;

(8) Rotate the condenser knob to move the condenser out of the optical path, and the field of view illumination is uniform.

5. Fluorescence observation:

(1) Place the fluorescent stained specimen on the stage.

(2) Transfer the 10X flat field objective or 40X fluorescent objective lens into the optical path, adjust the stage to move the handwheel vertically and horizontally, and move the specimen into the optical path.

(3) Rotate the filter conversion dial to transfer the excitation filter set required for the fluorescent staining specimen into the optical path. The excitation filter set number is engraved on the filter set change dial. Whether the filter is selected correctly or not is the key to the correct application of the fluorescence microscope. When selecting a filter, Stokes' law must be observed: the transmission wavelength of the excitation filter < the transmission wavelength of the two-color beam splitter < the transmission wavelength of the blocking filter.

Due to the excitation filter, the two-color beam splitter and the blocking filter, the factory has been strictly matched according to its application and its own optical characteristics. In observation and photography, you only need to select the filter group. .

(4) Adjust the coarse handwheel. After seeing the outline of the fluorescent image, adjust the focus with the fine adjustment handwheel until you see a clear fluorescent image.

(5) When a strong fluorescence image is needed, the condenser lens can be rotated to move the condenser into the optical path to obtain a brighter fluorescent image.

(6) When it is necessary to use a 40X or 100X fluorescent objective lens, glycerin should be added between the specimen and the objective lens, and there should be no vesicles or impurities in the oil that may affect the observation. When in use, the glycerin can be slowly infiltrated for a while, then gently rotate the animal mirror to remove the air bubbles.

6. Fluorescence microphotography - microscope camera

Since fluorescent images are generally much darker than bright field observations, it takes a long exposure time to perform fluorescence microscopy, and care should be taken to avoid instrument vibration during exposure.

Fluorescence microscopy is necessary for recording fluorescent images. Since fluorescence is easily faded, it is necessary to record the results in real time. The method is basically the same as ordinary microphotography.


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Address: No. 1271, Lincheng Street, High-tech Zone, Anshan City, Liaoning Province

Tel: +86 15642202798

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