Fluorescence microscopeUsed to study the absorption, transport, distribution, and localization of chemicals within cells. There are two forms of fluorescence produced by the measured object: self-luminescence. The fluorescence emitted directly after ultraviolet irradiation is called secondaryFluorescence。 After fluorescent dye treatment and ultraviolet irradiation, it can be emittedFluorescence。 Some substances in cells, such as chlorophyll, produce self-luminescence when exposed to ultraviolet light. Although other substances themselves do notFluorescence, but they can be used by fluorescent dyes or...FluorescenceAntibody staining is released twiceFluorescence。

A fluorescence microscope selects a high-efficiency point light source and emits light of a certain wavelength (ultraviolet 365nm or violet-blue light 420nm) as excitation light. It excites the fluorescent substances in the specimen, emitting semi-transparent light of various colors, which is then observed through the magnification of objectives and eyepieces. In such a strong contrast environment, even faint translucent light is easy to recognize and has very high sensitivity. It is mainly used to study cell structure, function, and chemical composition.

Fluorescence microscopeThey can be divided into two types: scattering microscopy and descent microscopy. The former is primitive, the latter is advanced. The basic structures of the two types of fluorescence microscopes are similar, with the main differences as follows: scattered excited light passes through the specimen, generating light throughout the specimen, which then enters the objective lens. The greater the magnification, the weaker the light; Incident excitation light is projected onto the sample surface, generating light on the sample surface, which then enters the objective lens. The greater the magnification, the higher the light exposure, making it suitable for high-magnification observation.

Fluorescence microscopyThe main components of the mirror include mercury lamp sources, excitation filters, color separation mirrors (for detachment), suppression filters, and dark-field spotlights (scattering). Additionally, due to the severe heating of mercury lamps, most are equipped with heat-absorbing filters. Some fluorescence microscopes also have phase difference mirrors and annular appendix, allowing observation of phase differences. For fluorescence microscopes, choose inverted frames, inverted microscopes, etc.

In addition, all the aforementioned microscopes have microscope CCDs assembled into digital microscopes, which will be...MicroscopeThe physical images seen are converted into digital images and displayed on a computer. Therefore, we can study micro-domains through traditional binocular observation and reproduction on displays, thereby improving work efficiency. 

ExistMetallographic microscopeDuring use, to observe metal damage, fractures must be smoothed, polished, and chemically washed before they can be seen.

1. Process: Simply put, they are divided into categories:Sampling→Inlay→Polishing and polishing→Erosion→Observation and photography are five steps.

II. Notes:

a) Metallographic specimens should only be gently ground on the side of the grinding wheel. When the thickness of the test piece is less than 10mm should be polished after inlay.

b) It is strictly forbidden to replace sandpaper or abrasive cloth while the grinding machine is running.

c ) When grinding and polishing, hold the test piece tightly and strive for smooth contact with the polished surface. Two people may not operate on the same spinning disk at the same time.

d) Chemical reagents for corrosive and electrolytic metallographic test strips should be stored and stored according to their properties, and relevant regulations must be followed during preparation and use; During electrolysis, the temperature and current density of the electrolyte should be strictly controlled.

e) The operating room for metallographic corrosion and electrolysis should be well-ventilated and equipped with tap water and solutions for emergency neutralization in case of acid or alkali injury.

f) Waste liquid used in metallographic tests must be treated as necessary before discharge; untreated waste must not be discharged into the sewer.

g) During on-site metallographic testing, measures should be taken to prevent reagents and solutions from spilling and dripping; After completion, all debris and waste liquids should be cleaned up.

h) Switch to a horizontal typeMetallographic microscopeWhen using the arc photoelectrodeYesCut off the power.

(1) When using an optical microscope, first check whether the overall installation of the microscope is loose, for example,Optical microscopeLong-term use can lead to problems such as loose support frames and overly loose lens installation. Too loose would harm the effectiveness of testing practical tests and easily damage the optical microscope.

(2) Common issues in using the focal screw setup. During operation, it is very easy to focus immediately under a high-magnification lens, or to keep both eyes looking at the eyepiece regardless of whether the barrel is raised or lowered, or to avoid the critical point of controlling the distance. When the distance is set to 2~3 cm, the focus increases further, and the rotation speed of the focus spiral is rapid. These problems easily lead to the objective lens lens being damaged during loading or damaging the mounting or camera lens. Regarding the above practical operation challenges, I want to say that when adjusting the lens focal length, you must land at low magnification. First, rotate the coarse focal length screw to gradually lower the barrel, bringing the objective lens close to the coverslip. Be careful not to let the objective touch the coverslip. Throughout this process, both eyes should look at the objective from the side, then use your right eye to look inside the eyepiece and slowly adjust the coarse focal length in the opposite direction so the barrel slowly rises until you see the virtual image. Also, note 'normal' for studentsOptical microscopeThe object distance is around 1 cm, so if the distance is well exceeded 1 cm but no virtual image is seen, this may be because the specimen collection is not within the field of view or the rotation of the rough focal screw is too fast. In this case, the loading area should be adjusted, and the process should be repeated. When a blurry virtual image appears in the field of view, a fine focal screw adjustment is required. Only in this way can the search range be reduced and the speed of virtual image search improved.

(3) The challenge of optical microscopes regarding light. Lighting is an applicationDuring optical microscopyA very important step, remember, is to use a low-power lens to direct the light. When the light source is strong, use a small focal length or a flat lens; when the light source is weak, use a large aperture lens or concave lens. When rotating the mirror, do not pull it out with one hand; rotate it with both hands until you see a uniform, bright circular field of view. After the light is properly aligned, never move the optical microscope arbitrarily, to prevent the light source from accurately entering the light source through the rearview mirror.

(4) The challenge of objective lens transformation. After using a low-power lens, switching to a high-power lens usually means people prefer to immediately rotate the objective lens with their fingers, thinking this saves effort. However, this easily causes the optical axis to tilt. The reason is that the original material of the converter is too soft in color, has high precision, and the external thread support is uneven, making it very easy for it to loosen. Once the external thread is damaged, the converter will be charged with the full inverter. Therefore, I recommend holding the next layer of the converter, the rotating plate to change the objective lens.

(5) High magnificationOptical microscopeThe challenge of using the eyes. When using a high-power microscope objective, the right eye should be as close as possible to the eyepiece. The right eye should try to look into the field of view, not forcefully covering the right eye or closing it decisively, as this violates experimental observation rules and often causes fatigue. Additionally, it cannot guarantee that the right eye is drawing while observing.

The condenser of a video microscope, also called a condenser, consists of a condenser and a variable aperturb. A condenser lens consists of one or more lenses, functioning like a convex lens, focusing light to enhance the light entering the objective. The variable aperture is also called the aperture, located below the condenser and consists of a dozen or so metal sheets, with a circular hole at the center. The handle that pushes the variable aperture can be freely adjusted to adjust the aperture size. Its function is to adjust the light intensity so that the numerical aperture of the condenser mirror matches that of the objective lens. The larger the variable light shutter opens, the larger the numerical aperture; Conversely, the numerical aperture becomes smaller.
The mirror of a video microscope is generally mounted on a base below the condenser and can rotate freely in both horizontal and vertical directions. One side is a flat mirror, and the other side is a concave mirror. Its function is to direct the light emitted from the source or natural light toward the condenser glass. Use a flat mirror when the light is strong, and a concave mirror when the light is weak. Some microscopes have artificial light sources installed inside the mount, and their mirrors are also fixed inside the frame.
A magnifying glass made of a single lens and a solid microscope composed of lenses (anatomical mirror) are called single-type microscopes. The optical microscopes used in biology teaching and research are generally compound microscopes.
Light incident from the outside is reflected upward by the mirror, or light emitted from the internal light source is viewed upward through the condenser mirror, then converged on the specimen under inspection by the condenser glass, providing sufficient illumination. Light reflected or refracted from the specimen enters through the objective lens, tilting the optical axis 45 degrees relative to the horizontal plane
A prism with a degree angle is magnified in the side light image at the focal plane of the eyepiece, that is, at the eyepiece's field of view light segment. This real image is then magnified into a virtual image through the eyepiece's eyepiece lens, so what people see is a virtual image.
The total magnification of the object under inspection after being magnified by the microscope's objective and eyepiece is the product of the objective lens and eyepiece. For example, the total magnification of a 40x magnified objective lens and a 10x magnified eyepiece is 400x.

When the magnification of a video microscope increases, on one hand, the higher the magnification, the more lenses there are, and the more light is absorbed by the lenses; On the other hand, since the brightness of the field of view (referring to the range of specimens being inspected) is inversely proportional to the square of the magnification, meaning the higher the magnification, the darker the field of view. To achieve sufficient brightness, a condenser must be installed to focus light on the specimen to be observed.
The condenser height of the video microscope at that point during observation
During observation, the best observation effect must be ensured; the condenser's focus should be exactly on the specimen. To achieve this condition, the height of the condenser must be adjusted. When illuminated with parallel light, the condenser's focus is about 1.25mm above the center of the upper lens plane. Therefore, it is often required to raise the condenser to the height of the stage plane just below the upper lens plane during observation, so the focus may fall on specimens on standard-thickness slides. When using slides thinner than the standard thickness to hold specimens, the position of the condenser should be lowered accordingly; when using slides that are too thick, the focus of the light only falls below the specimen, which is not conducive to detailed observation.
The condenser of a video microscope should be used together with the objective lens
The so-called coordination here refers to making the numerical apertures of the condenser and objective lens of the video microscope consistent, enabling more precise observation. If the condenser's numerical aperture is lower than the objective lens, part of the objective's numerical aperture is wasted and cannot achieve its maximum resolution. If the numerical aperture of the condenser is larger than that of the objective lens, on one hand, the objective's specified resolution cannot be improved; on the other hand, the illumination beam will be too wide, causing a decrease in image clarity. The method for working with the objective lens is: after completing illumination and focusing, remove the eyepiece and look directly into the tube, turn the variable light aphrodist under the condenser to the minimum, then slowly increase it. Open until its aperture is exactly the same diameter as the field of view, then put on the eyepiece to begin observation. Each objective switch requires a sequential coordination operation along with the transition. Some condensers have a scale indicating the opening aperture on the frame of the variable apertures, which can be adjusted according to the scale.

In microscope light source fault repairs, it was found that some users (about 25%) lack understanding of microscope light source systems and instead send microscopes that could have been resolved by themselves from far away or sent to their workplaces for repair. Not only is it time-consuming, but also labor-intensive and costly. To help these microscope users avoid unnecessary detours and unnecessary expenses, let me first introduce the repair methods when encountering a microscope light source failure:

1. Check if the bulb is burnt out. About 10% of users bring their microscopes to the manufacturer for repair because some filaments are burned and difficult to distinguish with the naked eye.
2. Check if the fuse is blown. About 5% of users return to repair because some fuses are difficult to visually identify after blown-out fuses. The correct way to check is to use a multimeter in the resistance range to check if the fuse is in good condition
3. Check whether the microscope bulb socket has poor contact due to surface oxidation due to high temperature. About 7% of users bring the microscope for repair due to oxidation on the lamp socket surface. The correct inspection method is to remove the bulb, use fine sandpaper or a small blade to sand off the oxide layer on the lamp holder surface, then reinstall the lamp to check if the socket has good contact.
4. Check whether the power socket is making good contact. About 3% of users bring microscopes with no light source issues to the manufacturer for repair, but after plugging in the power, the microscope's light source works normally.

When using a microscope, the first thing to consider is what you need to observe and what kind of observation/imaging results you want to achieve, before deciding which type of microscope to use. If the ideal clear-field microscope effect is to be achieved (referring to the microscope's optimal resolution), but the microscope uses achromatic or phase-difference objectives, or the aperture of the condenser is only 1.2 (Abbe condenser), then no matter how hard you try, it is impossible to take the best photo (when working with an oil filter).

Therefore, before shooting, it is crucial to check whether the type and components of the microscope used are compatible, and whether the adjustment method is correct. Generally, to determine whether the microscope method is correct, first check the type of objective, then check whether the condenser is used correctly, and finally verify whether it meets Kohler illumination requirements.

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