Anatomy of a DSLR - dslr anatomy

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(2006). Multiphoton Excitation and Microscopy. In: Lakowicz, J.R. (eds) Principles of Fluorescence Spectroscopy. Springer, Boston, MA. https://doi.org/10.1007/978-0-387-46312-4_18

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Until 1990 multiphoton spectroscopy was considered to be an exotic phenomenon that was used primarily in chemical physics and optical spectroscopy. Two-photon ab-sorbance or excitation requires high peak powers to increase the probability that two photons are simultaneously available for absorption. Because of the interaction of two photons with the fluorophore, the selection rules for light absorption are, in principle, different from those for one-photon spectroscopy. Because of the different selection rules, two-photon spectroscopy can be used as a tool to study the excited-state symmetry of organic chro-mophores.1–3 Multiphoton experiments require complex lasers and high optical powers. It did not seem possible to use multiphoton excitation (MPE) in optical microscopy because the high power would damage the biological samples. Surprisingly, MPE is now widely used in fluorescence microscopy. Multiphoton microscopy (MPM) is possible because of the favorable properties of titanium—sapphire (Ti:sapphire) lasers and the development of laser-scanning microscopes. Multiphoton excitation is usually less damaging to biological samples than in one-photon excitation. Multiphoton microscopy was introduced in 19904 and is now used extensively in cell imaging.5–9

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Crystalline material UV Band Pass Filters (available crystalline materials - NiSO4*6H2O (NSH) and K2Ni(SO4)2*6H2O (KNSH)), neutral density reflective type filters made from Corning 7059 glass and designed to operate at 400-2000 nm range, neutral density absorption type filters designed to operate at VIS (450-650 nm) and Schott color glass filters.

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25. Lakowicz JR, Gryczynski I. 1992. Fluorescence intensity and anisotropy decay of the 4′,6-diamidino-2-phenylindole–DNA complex resulting from one-photon and two-photon excitation. J Fluoresc 2(2):117–122.

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In the previous chapters of this book we described the emission resulting from one-photon excitation (1PE). By 1PE we mean that an excited fluorophore has reached the excited state by absorption of a single photon. We now consider two-photon (2PE) and three-photon (3PE) excitation. The term 2PE indicates that the fluorophore has reached the excited state by absorption of two photons. We will only consider simultaneous absorption of two or more photons. We will not consider sequential absorption where there is a well-defined intermediate state.

Optical filter

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