Fluorescence microscope is immune cells of fluorescent chemical basic tools. It is by the light source, filter board system and optical system and other major parts. Is the use of certain wavelengths stimulate specimens launch fluorescence, through the optical lens and eyepiece system enlarged to observe samples of fluorescence image

(a) light source

Now with 200 W of ultra high pressure more mercury lamp as the light source, it is used with quartz glass production, and the middle is spherical, filling in a certain number of mercury, work by the two terminals discharge, cause mercury evaporation, ball pressure within lifts quickly, when mercury completely evaporation, can be up to 50 ~ 70 standard atmospheric pressure, this process generally need about 5 to 15 min. The shine is super high pressure mercury lamp between electrodes that mercury molecular continuous discharge disintegrate and reduction process launch the result of light quantum. It launch strong ultraviolet and LanZiGuang, enough to inspire all kinds of fluorescent material, therefore, widely used for a fluorescent microscope.

High pressure mercury lamp also sends out a lot heat energy. Therefore, the light room must be good thermal conditions, is not too high temperature environment.

New super high pressure mercury lamp in the early use need not be high voltage can be ignites, use some time, they need to start high pressure (about 15000 V), start, maintain working voltage for 50 to 60 V general, working current about 4 A or so. 200 W of ultra high pressure mercury lamp average life expectancy, at every time of 2 h, use about 200 h, start a shorter work, then the shorter life expectancy, such as having a work only 20 min, life is reduced by 50%. Therefore, when using, try to reduce the start times. The light bulb in use process, its efficacy is gradually decreasing. After the lights went out to wait for cooling to restart. Light bulb not immediately after the close, in order to avoid mercury evaporation completely and damage the electrode, generally need to wait for 15 min. Due to the high pressure mercury lamp pressure is high, ultraviolet ray strong, so the light bulb must buy lamp lit room the Chinese party may, in order to avoid damage eyes and explosion caused when the operation.

High pressure mercury lamp (100 W or 200 W) light source circuit and including variable pressure, ZhenLiu, start, several parts. In the light bulb glow has adjust the room in the center of the system, the light bulb behind the ball to install coating aluminum of concave mirrors, are installed in front of the lens with light.

Domestic ultrahigh pressure mercury lamp GCQ-200 type performance is good, can take the place of the HBO-200 type such as the import of the light bulb, average life expectancy in more than 200 h, prices are low.

Our country developed a simple light model high color temperature fluorescent light source device bromine tungsten, small volume, light weight, small power, ac, dc amphibious (bring dc power), easy to carry, easy to use, has been applied.

(2) color filters system

Color filters system is an important part of the fluorescence microscope, stimulate filter board and suppressed by the filter board composition. Filter board type, each manufacturer are often not unified. Filter board usually with basic tonal named, front letter stands for tonal, behind letter stands for glass, number represents model characteristics. Such as German products (Schott) BG12, is kind of blue glass, B is the first letter of the blue, G is the first letter of the glass; The name of the product in China have agreed to use pinyin letter said, such as equivalent to BG12 blue filter board called QB24, Q is blue (blue) the first letter of the alphabet, B is glass the first letter of the alphabet. But some filter board can also pervious to light boundary named long filter, such as K530, is suppressed long 530 nm filter said the light and through more than 530 nm of light. And the manufacturer filter board completely with digital naming, such as the United States Corning factory of NO: 5-58, that is BG12.

1. According to stimulate filter board light source and the characteristics of the fluorescent pigments, can choose the following three stimulate filter board, provide a certain wavelength range stimulate the light.

Ultraviolet stimulate filter board: the filter board can make the 400 nm the following through the ultraviolet light, blocking the visible through more than 400 nm. Commonly used models for UG-1 or UG-5, plus a BG-38, to remove red tail wave.

Ultraviolet blu-ray stimulate filter board: the filter board can make 300 ~ 450 nm range of light through the. Commonly used models for ZB-2 or 3 ZB, plus BG-38.

Violet blue stimulate filter board: it can make 350 ~ 490 nm light through the. Commonly used models for QB24 (BG12).

Maximum absorption peaks in the 500 nm older fluorescent element (such as Rhoda Ming pigment) available blue green filter board (such as B-7) motivated.

In recent years, beginning to use a metal interference filter board, because the targeted, appropriate wavelength, and stimulate the effect is better glass filter. Such as west Germany Leitz factory of special KP490 FITC filter board and Rhoda Ming S546 green filter board, are far more than glass filter board the effect is good.

Stimulate filter board points bao hou two kinds, general dark field choose thin filter board, bright vision fluorescence microscope can choose thicker. Basic requirements is the most bright fluorescent and get the best background shall prevail.

2. Suppress filter board suppress filter board's role is completely block stimulate light through, to provide corresponding filter long range fluorescence. And stimulate the filter board corresponds, commonly used the following 3 kinds of pressing filter board:

Ultraviolet suppress filter board: through the visible light, stop by ultraviolet light. Can and UG-1 or UG-5 combination. Commonly used GG-3 K430 or GG-6 K460.

Violet blue suppress filter board: can through the 510 nm filter over long fluorescent (green to red), can and BG-12 combination. Usually use OG-4 K510 or OG-1 K530.

Uv lights suppress filter board: can through the 460 nm wavelength above the fluorescent (blue or red), but with BG-3 combination, commonly used OG-11 K470AK490, K510.

(3) reflector

The reflective layer must is generally aluminium, for aluminum to uv light and visible light blue purple area absorb less, the reflection of 90% above, and the reflex of the silver only 70%; General use plane reflector.

4) condenser

Designed for a fluorescent microscope design manufacture parabolic dish concentrator is with quartz glass or other through made of glass, uv light. Clear vision parabolic dish concentrator's dark field parabolic dish concentrator two kinds. And differ fluorescence parabolic dish concentrator.

1. The vision parabolic dish concentrator fluorescence microscope in general on the use of Ming vision parabolic dish concentrator, it has contracts to force strong, easy to use, particularly suitable for low, in times of amplification specimen observation.

2. Dark field parabolic dish concentrator dark field parabolic dish concentrator in a fluorescent microscope has been used increasingly in. Because inspire not directly into the objective light, so in addition to the scattering outside, stimulate the light also not entering the eyepiece, can use thin stimulate filter board, enhance stimulate the strength of the light, the suppression of filter board can also be very thin, because ultraviolet stimulate, usable colorless filter board (not through ultraviolet) and still produce dark background. And improve the brightness and contrast of fluorescent image degree, improve the quality of the image, observation and comfortable, may find bright vision of difficult to recognize subtle fluorescent particles.

3. Differ fluorescence parabolic dish concentrator are parabolic dish concentrator is used together with the objective, but at the same time difference and fluorescence joint observation, already can see fluorescence image, and can see the difference image, help to fluorescent accurate location. General fluorescent observation rarely need this kind of parabolic dish concentrator.

(5) the objective

The objective of all can be used, but the best is away of these color difference, because of its auto-fluorescence microscopic and pervious to light performance (the wavelength range) suitable for fluorescence. Due to the perspective of fluorescence microscope images in brightness and the objective lens is proportional to the mouth of the rate, and inversely proportional to magnification, so in order to improve the image of the fluorescence intensity, should use rate of these big mouth lens. Especially in the influence at high magnification is very obvious. So on fluorescence of the specimen is not strong, should use rate of these big mouth lens, cooperate with the lowest possible eyepiece (4 x, 5 ×, 6.3 by, etc.).

(6) the eyepiece

In the use of low power eyepiece fluorescence microscope, such as a 5 × and 6.3 x. The past is multi-purpose list tube eyepiece, because its brightness than double barrel twice as tall as the eyepiece above, but the present research fluorescence microscope multi-purpose eyepiece double tube, observation is very convenient.

(7) fall shot all devices

The new fall shot all device is the light of light sources to shoot to interfere in spectral filters, wavelength a short part (ultraviolet and violet blue) due to the nature of the coating on the filter and reflection, when filter to light source of a 45. Tilt, the vertical into the objective, the optical lens into specimens, make specimen is motivated, then directly on the objective of parabolic dish concentrator role. At the same time, the filter long part (green, yellow, red, etc), to filter through is, therefore, not the direction to the objective reflection, filters, plays a triggering filter board effect, because of the specimen in visible light long fluorescent area, can through to filter the eyepiece observation, the brightness of the fluorescent image magnification increase and improve with, in high amplification than transmission light source is strong. It is divided outside the function of transmission type illuminant, more suitable to the opaque and translucent specimens, such as thick slices, the membrane filter, colonies, tissue culture of specimens, directly observed. In recent years the development of new fluorescent microscopy used more fall shot all device, called the fall shot a fluorescent microscope.