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Quantitative measurement of the resolution and sensitivity of confocal microscopes using line‐scanning fluorescence correlation spectroscopy
Author(s) -
Balaji J.,
Maiti S.
Publication year - 2005
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20162
Subject(s) - confocal , microscope , fluorophore , resolution (logic) , confocal microscopy , fluorescence correlation spectroscopy , 4pi microscope , microscopy , optics , spectroscopy , materials science , measure (data warehouse) , sensitivity (control systems) , fluorescence microscope , fluorescence , physics , computer science , artificial intelligence , data mining , multiphoton fluorescence microscope , quantum mechanics , electronic engineering , engineering
Abstract Spatial resolution and the sensitivity to detect a fluorophore are the two most important optical parameters that characterize a confocal microscope. However, these are rather difficult to estimate quantitatively. We show that fluorescence correlation spectroscopy (FCS) provides an easy and reliable measure of these quantities. We modify existing schemes for performing FCS on a commercial confocal microscope to carry out these measurements, and provide an analysis routine that can yield the relevant quantities. Our method does not require any modification of the confocal microscope, yet it yields a robust measure of the resolution and sensitivity of the instrument. Microsc. Res. Tech. 66:198–202, 2005. © 2005 Wiley‐Liss, Inc.