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2-PhoExcMic-2011

2-PhoExcMic-2011 - TWO PHOTON EXCITATION MICROSCOPY 1931...

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TWO PHOTON EXCITATION TWO PHOTON EXCITATION MICROSCOPY MICROSCOPY
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HISTORY OF TWO HISTORY OF TWO - - PHOTON PHOTON EXCITATION MICROSCOPY EXCITATION MICROSCOPY 1931: The concept first described by Maria Goppert-Mayer in her doctoral dissertation. Observed experimentally some thirty years later, shortly after the invention of the laser. It was pioneered by Winfried Denk at Cornell University.
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WHY TWO WHY TWO - - PHOTON EXCITATION PHOTON EXCITATION MICROSCOPY ??? MICROSCOPY ??? It provides distinct advantages for three-dimensional imaging. It excels at imaging of living cells, especially within intact tissues such as brain slices, embryos, whole organs, and even entire animals .
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Principles of Two Principles of Two - - Photon Excitation Photon Excitation Two-photon excitation arises from the simultaneous absorption of two photons in a single quantitized event. Since the energy of a photon is inversely proportional to its wavelength, the two absorbed photons must have a wavelength which is about twice that required for single photon excitation, i.e, a fluorophore that normally absorbs UV at 350nm can alternatively be excited by 2 red photon at 700nm provided they excite the fluorophore at the same time (~10 -18 sec) The fluorescence emission depends on the square of the excitation intensities.
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Principles of Two Principles of Two - - Photon Excitation Photon Excitation (Contd.) (Contd.) (1) (1)
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Two Two - - Photon Excitation in Laser Photon Excitation in Laser - - Scanning Scanning Microscopy Microscopy Based on the principle that absorption depends on the square of excitation intensity.
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