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Advantages Of Two-photon Microscopy Over Confocal Microscopy
Release time:
2024-08-21
The Principle Of Two-photon Absorption/excitation Technology In Two-photon Microscopy: Under Strong Light Excitation, Molecules Simultaneously Absorb Two Photons And Transition From The Ground State To An Excited State With Twice The Energy Of The Photons. Two Photons Can Be Of The Same Wavelength Or Different Wavelengths, But They Must Be Absorbed Simultaneously (the Time Interval Between The Two Photons Reaching The Excited Molecule Is Less Than 1 Femtosecond). It Can Be Understood As Absorbing The Energy Of A Photon First To Transition To A Virtual Intermediate State, And Then Absorbing The Energy Of A Photon To Transition To An Excited State.
Advantages Of Two-photon Microscopy Over Confocal Microscopy And Other Microscopes:
Low Light Damage In Two-photon Microscopy: Due To The Use Of Visible Or Near-infrared Light As The Excitation Light Source, This Wavelength Of Light Causes Minimal Light Damage To Living Cells And Tissues, Making It Suitable For Long-term Research;
Strong Penetration Ability Of Two-photon Microscope: Compared With Ultraviolet Light, Visible Light And Near-infrared Light Have Stronger Penetration Ability, So They Are Less Affected By The Scattering Of Biological Tissues, Solving The Problem Of Tomographic Imaging Of Deep Substances In Biological Tissues; Research Has Shown That The Imaging Depth Of Confocal Fluorescence Imaging Is Generally Around 50 Microns, While The Imaging Depth Of Two-photon Microscopy Can Reach Up To 1000 Microns;
High Resolution Two-photon Microscope: Due To The Small Absorption Cross Section Of Two-photon, Fluorescence Can Only Be Excited In A Very Small Area Of The Focal Plane, And Two-photon Absorption Is Limited To The Volume At The Focal Point, Which Is About The Third Power Of The Wavelength.
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