Introducing the advantages and disadvantages of multiphoton laser scanning microscopy
Advantages of multi-photon laser scanning microscope:
1, using red or infrared light excitation, light scattering is small, the scattering of small particles is inversely proportional to the fourth power of the wavelength.
2,No pinhole is needed to collect more scattered photons from the imaging cross section.
3, the pinhole can not distinguish between the scattered photons emitted by the out-of-focus region or focal area, multi-photon in the deep imaging signal-to-noise ratio is good.
4, single-photon excitation of ultraviolet or visible light used in the beam to reach the focal plane before the sample is easily absorbed and attenuated, not easy to deep excitation.
5, in the biological microscope observation, * the first consideration is not to damage the active state of the organism itself, to maintain moisture, ion concentration, oxygen and nutrient circulation. In light observation occasions, both thermal and photon energy must stay in the cell without damage to the amount of irradiation, light energy.
6, multi-photon microscope has many advantages. Such as three-dimensional resolution, depth intrusion, in the scattering efficiency, background light, signal-to-noise ratio, control, etc., have the previous laser microscope does not have or can not be compared to surpass the characteristics of the
Disadvantages of multi-photon laser scanning microscope:
1, only fluorescence imaging.
2, if the sample includes chromophores that can absorb the excitation light, the sample may be thermally damaged.
3, the resolution is slightly reduced, although it can be improved by the use of confocal aperture at the same time, but the signal loss.
4. The cost of multiphoton scanning microscopy is high due to the limitation of expensive ultrafast lasers.
