Evanescent field excitation of fluorescence by epi-illumination microscopy Export

@article{citeulike:5141513, abstract = {By simple modification of the pattern of fluorescence excitation light in an epi-illumination inverted microscope, one can achieve conditions that produce total internal reflection fluorescence (TIRF) by evanescent wave excitation. Though traditionally requiring a collimated beam traversing through a special prism, TIRF also can be achieved by epi-illumination through the periphery of a 1.4 numerical aperture objective. An opaque disk of appropriate size is placed in the illumination path external to the microscope so as to cast a sharp, real-image shadow at the objective's back focal plane. This shadow allows a hollow cone of epiillumination rays traveling at only super-critical angles to reach the glass/water interface at the sample plane. Three kinds of TIRF illumination patterns can be produced by variations of this scheme: (1) a small spot of illumination of 1.5 µm radius by use of a laser light source, (2) a large region of illumination by use of a laser-illuminated diffusing screen located upbeam from the opaque disk, and (3) a large region of illumination by use of a conventional mercury arc.}, author = {Stout, Andrea L. and Axelrod, Daniel}, citeulike-article-id = {5141513}, citeulike-linkout-0 = {http://dx.doi.org/10.1364/AO.28.005237}, citeulike-linkout-1 = {http://www.opticsinfobase.org/abstract.cfm?id=33056}, day = {15}, doi = {10.1364/AO.28.005237}, journal = {Appl. Opt.}, keywords = {history, microscopy}, month = {December}, number = {24}, pages = {5237--5242}, posted-at = {2009-07-14 01:37:19}, priority = {2}, publisher = {OSA}, title = {Evanescent field excitation of fluorescence by epi-illumination microscopy}, url = {http://dx.doi.org/10.1364/AO.28.005237}, volume = {28}, year = {1989} }

TY - JOUR ID - citeulike:5141513 L3 - citeulike-article-id:5141513 N2 - By simple modification of the pattern of fluorescence excitation light in an epi-illumination inverted microscope, one can achieve conditions that produce total internal reflection fluorescence (TIRF) by evanescent wave excitation. Though traditionally requiring a collimated beam traversing through a special prism, TIRF also can be achieved by epi-illumination through the periphery of a 1.4 numerical aperture objective. An opaque disk of appropriate size is placed in the illumination path external to the microscope so as to cast a sharp, real-image shadow at the objective’s back focal plane. This shadow allows a hollow cone of epiillumination rays traveling at only super-critical angles to reach the glass/water interface at the sample plane. Three kinds of TIRF illumination patterns can be produced by variations of this scheme: (1) a small spot of illumination of 1.5 µm radius by use of a laser light source, (2) a large region of illumination by use of a laser-illuminated diffusing screen located upbeam from the opaque disk, and (3) a large region of illumination by use of a conventional mercury arc. IS - 24 JF - Appl. Opt. EP - 5242 TI - Evanescent field excitation of fluorescence by epi-illumination microscopy PB - OSA VL - 28 SP - 5237 KW - history KW - microscopy AU - Stout, Andrea AU - Axelrod, Daniel PY - 1989/12/15/ UR - http://dx.doi.org/10.1364/AO.28.005237 ER -