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Stereo Microscope Correct Method And Method Of Use

After understanding the name, structure and function of the main components of the Stereo Microscope, in order to better play the various functions of the Stereo Microscope, improve work efficiency, to ensure that the microscopic observation and micro-photographic process to achieve the best results, the use of personnel must Understand and master the correct debugging method and use of the Stereo Microscope. Especially in the new generation of Stereo Microscopes, with a variety of functions, can carry out a variety of microscopic observation method, the correct test method and the use of methods is particularly important. The following to universal research Stereo Microscope, for example, a brief description of debugging and use.

1. Stereo Microscope lighting optical system adjustment

In order to make the Stereo Microscope field of view can be uniform and full of lighting, the initial installation and commissioning of the Stereo Microscope, it is necessary to adjust the lighting optical system, which is the correct use of the Stereo Microscope, and get the correct, * results of the important means and the most basic The request. In addition, the correct control of the lighting system of light adjustment system is to use the Stereo Microscope to replace the light bulb after the necessary steps, but also in the daily use of the Stereo Microscope from time to time to test the performance of the necessary means. Stereo Microscope lighting optical system adjustment mainly has the following four elements:

(1) lighting the light room in the Stereo Microscope outside the initial adjustment

(1) Open the housing of the lamp compartment first, press the spring bulb into the socket, install the ground to avoid direct contact with the lamp (soft cloth or paper can be separated), so as not to leave fingerprints and other dirt, affecting the bulb Of the service life.

② the lamp room on the table, turn on the power, with a special screwdriver to adjust the lamp knob knob (marked "← →"), the filament projection in the 1-2m outside the wall, the filament imaging To the clear; and then adjust the height of the adjustment of the lamp position adjustment hole (marked "-"), so that the filament position high and low appropriate; and then adjust the left and right position adjustment screw hole (marked "-"), so that the filament left and right position The

(2) light source light (filament) in the Stereo Microscope within the location of the inspection and correction

The purpose is to keep the luminous body from the perspective of the light source to ensure that the Stereo Microscope's field of view is fully and evenly illuminated, which is a prerequisite for adjusting the Kule lighting system. The basic tools needed are: when the telescope is equipped with a Stereo Microscope.

① unplug the light glass inside the lamp sleeve, the lamp room back to the Stereo Microscope;

② use 10 × objective lens, open the light source program to find samples and focus clear, and then use 40 × objective lens to focus on the sample clear (40 × objective lens can see the whole picture)

③ the condenser aperture aperture and the aperture of the aperture are open to the maximum;

④ unplug one of the eyepiece, put on the telescope, seize the white part, the other hand telescopic black eyepiece, you can see in the field of light filament;

⑤ If the filament position is not appropriate, adjust the "hole", the filament along the horizontal direction of tune, tune "─ ─" hole, the silk along the vertical direction, until the filament is just like the lens aperture Photosphere

⑥ after adjustment, the hair glass sleeve into the original position, unplug the telescope, in exchange for the next step to adjust the eyepiece. The above adjustment of the lighting room of the illumination light source outside the Stereo Microscope and the verification of the position of the light source in the Stereo Microscope are carried out only when the Stereo Microscope is first installed and commissioned and the lamp is replaced. The Stereo Microscope can not be used when the Stereo Microscope is used. In case of disruption, according to the above steps back to the original state.

(3) the correct adjustment of the Kohler system

One of the main work of the correct debugging of the Stereo Microscope is the adjustment of the lighting system, and the key is the adjustment of the Kule lighting system. For each person who uses the Stereo Microscope, especially for photographed personnel, should be on the Kule lighting system principle and its adjustment steps have a certain understanding and master, in order to give full play to the function of the Stereo Microscope, Out of the photos in order to be more consistent and perfect. The principle of the Kule lighting system is simply that the light emitted from any point on the light source illuminates the range of the Stereo Microscope, and the light emitted by each point on the light source emits together, and in the field of view of the Stereo Microscope To achieve a very full and uniform lighting. The purpose of the Korla lighting system is to allow uniform and adequate illumination of the observed field of view to prevent stray light from affecting or interfering with the system, so as to avoid fogging in the film when photographed. Highly adjustable Kule lighting system necessary components: field diaphragm, can be adjusted to adjust the condenser system.

① use 10 × objective lens and 10 × eyepiece;

② the condenser lens into the front of the optical path, the aperture aperture to a moderate position (not much), then the condenser to the top of the position, condenser mirror turn to the bright field "J" position;

③ adjust the field of view to the smallest aperture (0.1);

④ put on the stage of the biological samples have been sealed, open light source, focusing clear;

⑤ There will be a localized area or bright spot in the field of view, which is a blurred image of the field of view, where the details of the sample can be clearly seen; it is a darker view outside it. The details of the sample see clearly;

⑥ to condenser slightly down, so that the field of light in the bright spot gradually small, slowly into a clear multilateral image, which is a clear vision of the aperture;

⑦ Under normal circumstances, the multilateral image is not in the center of the field of vision, you need to adjust the condenser of a pair of adjustable screws, the aperture of the aperture diaphragm as the central position;

⑧ gradually open large field aperture, so that the multilateral image into the field of the internal polygon, further check the tone of the situation, such as the middle is not ideal, continue to fine on the screw;

⑨ the aperture of the aperture slightly to open slightly larger, so that its multilateral image just disappeared in the sight of the edge, thus, Kule lighting system adjustment is completed. Kule lighting system adjusted well after the entire field of view lighting uniform, photographed photomicrograph bright and clear, contrast normal. In the future use of the process should pay special attention to: a. Field diaphragm can not be arbitrarily opened, but with the increase in the magnification of the lens and the aperture will be small, with the reduction of the objective lens to open large; b. Condenser C) the use of 10 × below the objective lens to the front end of the condenser lens placed outside the optical path, the use of 10 × or more than 10 × when the objective lens to the front end of the lens The lens into the light path; d. On the objective lens multiplier and the aperture of the size of the problem, in the actual use of the process, as a general observation does not have to close or open large field diaphragm, but for micro photography, in order to To avoid the interference of the stray light to the camera system in order to be able to take a better picture, you should use the objective lens of each multiples to adjust the field diaphragm just to the side of the observed field. More complicated work, but not do not. The simpler method is to adjust the field of view corresponding to each objective lens, and make a mark, and use it directly to the corresponding position according to the mark.

(4) the correct use of aperture stop

As the aperture of the aperture aperture diaphragm can affect the resolution of the Stereo Microscope, the use should be the correct use of the method. In the past, due to lack of understanding of the aperture stop, it is often regarded as a tool to adjust the brightness of the field of view. Although the aperture aperture can be adjusted to a certain extent, can change the brightness of the field of vision, but will directly affect the imaging contrast, contrast and resolution, the use of the process should be avoided. In order to achieve the role of the condenser aperture aperture, in order to observe, especially in the micro-photography to obtain the best resolution, each in a multiples of the objective lens, the sample focus is clear, the need to adjust the aperture stop, So that its size is exactly equal to the use of objective lens aperture (objective lens aperture) 2/3. The adjustment method is to focus on the telescope in the field of view on the black phase difference, adjust the aperture stop, you can see a polygon aperture aperture, and then transferred to equal to the objective lens aperture like 2/3, that is, between the black phase difference ring Between the outer and the circular view. For the sake of convenience, the aperture stop corresponding to each objective lens can be pre-adjusted and marked so as to be readjusted every time it is used.

2. Stereo Microscope imaging optical system adjustment and microscopic examination outline

Stereo Microscope imaging optical system adjustment, is based on the needs of different microscopic examination carried out. The so-called microscopy (microscopy), in general, is a Stereo Microscope to observe the sample when used in the lighting method, and how to make the sample into the image can get better contrast technology and methods. The following is a brief description of the microscopic examination has been mature in several ways and the corresponding Stereo Microscope imaging optical system adjustment method.

(1) Transparent light field of vision

This is the most traditional and popular method since the invention of the Stereo Microscope. Basic components: a. Objective: any objective lens can be used for visual field observation; b. Condenser: a variety of condenser can be, preferably equipped with aperture stop. Adjustment method: in the Stereo Microscope Kule lighting system is adjusted, you can apply the bright field method. Scope: All stained tissue sections, blood smears and so on. Note: a. Use the visual field method of observation, we must adjust the Kule lighting system; b. Field diaphragm can not be arbitrarily open, the use of 10 ×, 10 × and 10 × above the objective lens, the condenser C. Can not use the aperture of the aperture aperture to adjust the brightness of the field of view, not to blur the high and low position of the condenser, otherwise, will reduce the resolution and damage of the Stereo Microscope has been adjusted good Kule lighting system; d. For micro-photography, each for a multiple of the objective lens, it is necessary to adjust the condenser aperture aperture, so that its size is exactly equal to the objective lens diameter of 2/3.

(2) transmitted light phase difference method

This is a contrast enhancement in modern microscopy. Basic components: the difference between the objective lens, bright field and the difference between the multi-purpose condenser, the binoculars, green filters. Adjust the method: a. In the Kule lighting system on the basis of a good adjustment, with the bright field method to focus the sample clear; b. The condenser to Ph1 alignment dial scale line position, the choice of 10 × phase difference lens, put on to be observed Of the transparent sample; c. Unplug one of the eyepieces, put on the telescope, and focus on the field of view of the two phase difference (the objective lens of the black phase difference and condenser light transmission phase difference); d. The two phase difference rings do not coincide with each other, adjust the two adjustment devices on the condenser lens (adjust the adjustment lever of the position difference between the left and right position and adjust the front and rear position of the friction type knob), so that the light transmission ring around the black and the left and right; E. After adjustment, change the observation eyepiece, the green filter into the light path, you can observe the sample phase difference; f. 20 × and 40 × objective lens observation, the condenser should be located in the Ph2 position , With 100 objective lens, the condenser should be located in Ph3 position. Scope of application: Suitable for observing transparent, unstained or non-dyed samples, such as various cells, living tissue, unstained or non-stained tissue sections, aquatic organisms.

(3) differential interference phase contrast method

In order to overcome the difference between the observation of the sample details like the surrounding with a halo, will cover up the details should have been seen, and the sample or tissue slice thickness requirements are quite thin, the principle can be thicker than 10? M and other limitations, the use of dual-beam Principle of interference design sub - differential interference phase contrast method. Adjust the method a. Must be in the Kulaiming system has been adjusted on the basis of the DIC method can be adjusted; b. First with 10 × objective lens to clear the first look at the sample can be clear to see the objective lens focus position; The polarizer is placed in the illumination light path and the orientation should be in the east-west direction; d. Turn the condenser wheel to the position corresponding to the 10 × objective lens, ie DIC 0.3-0.4; e. Or the DIC slider used to insert the 10x objective lens on the objective converter; f. Insert the analyzer into the imaging light path, note that the orientation should be south-north; g. Replace the transparent Sample, open the light source to focus the sample clear; h. Adjust the DIC insert, so that the differential interference phase contrast to achieve the best results, that is, the most obvious relief effect; i. At the same time adjustable condenser aperture aperture, The effect is also the best; j. And then fine-tune the details of the sample, we can see the structure of the different layers of the sample; k. If the first order red retardation plate inserted, and adjust the DIC insert, Look in the field of view Changing brilliant colors, red, orange, yellow, green, blue, purple, pink, purple and gold have. Scope: transparent or can not be stained tissue sections, the thickness of up to 100? M or so, the culture of living tissue and living cells, niches and so on.

(4) Fluorescence excitation of emission light

Referred to as the ablation fluorescence method, is a modern Stereo Microscope in the new development of a strong contrast enhancement method. It will stimulate the fluorescent light source to change above the objective lens, the light from the top of the objective lens through the mirror into the objective lens to stimulate the sample, from the sample to stimulate the fluorescence through the objective lens and penetrate the mirror and observed by the eyepiece. The method is simpler and more efficient, and the intensity of the 50W light source is stronger than the 250W of the transmission fluorescence method. Fluorescence method is to use the wavelength of short ultraviolet light, violet, blue violet, blue and green light to stimulate the sample, as long as the sample contains a fluorescent component, it absorbs short-wave excitation light and the release of longer wavelength fluorescence The Different substances can only absorb specific wavelengths of excitation light, and the release of fluorescence will have a specific wavelength, and thus for the identification of specificity is very effective, such as certain pathogenic bacteria and spirochetes, by ultraviolet light can be issued after excitation Their unique fluorescence, it is easy to make identification, the use of substances to absorb the excitation of light after the release of specific fluorescence method known as the spontaneous fluorescence method. Some substances themselves do not absorb the excitation light, or can not absorb the fluorescence after absorption, but can absorb or absorb specific fluorescent pigments or dyes, and these specific fluorescent pigments or dyes can only absorb specific excitation light, and then release a specific Of the fluorescence, which indirectly identify a substance, which is called indirect fluorescence method. The fluorescence method is widely used in medical, biological and industrial specific research and identification. Adjustment method: fluorescence Stereo Microscope or Stereo Microscope with fluorescent parts, the adjustment method is roughly the same.

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