(Differences between (two-photon, confocal) fluorescence microscopes and ordinary microscopes
Fluorescence microscope is the use of ultraviolet light as a light source, used to irradiate the object to be examined, so that the object emits a light source, and then the observation of the object is carried out under the microscope. It is mainly used for immunofluorescence cells, which is mainly composed of light source, filter plate system and optical system to observe the fluorescence image of the samples through the magnification of eyepiece and objective lens. Let's look at the fluorescence microscope and ordinary optical microscope has what kind of difference.
1, in the illumination mode
The illumination of fluorescence microscope is generally with the falling type, that is to say, the light source is through the objective lens to put on the test sample.
2, in the resolution
Fluorescence microscope uses ultraviolet light as a light source, the wavelength is relatively short, but the resolution is higher than ordinary optical microscope.
3, the difference in the filter
Fluorescence microscope is the use of two special filters, in front of the light source is used to filter out the visible light, between the objective lens and the eyepiece to filter out the ultraviolet light, which can protect the human eye.
Fluorescence microscope also belongs to a kind of optical microscope, mainly fluorescence microscope excitation wavelength is short, so this led to the fluorescence microscope and ordinary microscope miscellaneous structure construction and use of different, fluorescence microscope most have good capture of weak light function, so in the extremely weak fluorescence, it's imaging ability and good. Coupled with the continuous improvement of the fluorescence microscope in recent years, the noise is also greatly reduced. Therefore, more and more fluorescence microscopes have been applied.
Two-photon fluorescence microscope has many advantages:
(1) Longer wavelengths of light are less affected by scattering than shorter wavelengths of light easily penetrate the specimen;
(2) The fluorescent molecules outside the focal plane are not excited so that more excitation light can reach the focal plane, so that the excitation light can penetrate deeper into the specimen;
3) Longer wavelengths of near-infrared light are less toxic to cells than shorter wavelengths;
4) When using a two-photon microscope to observe a specimen, photobleaching and phototoxicity are present only at the focal plane. Therefore, two-photon microscopes are more suitable than single-photon microscopes for observing thick specimens, for observing living cells, or for conducting spot photobleaching experiments.
