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Proceedings Paper

Using an adjustable detection time to correct photobleaching effects in fluorescence microscopy
Author(s): Eric W. Hansen
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Paper Abstract

Photobleaching causes progressive fading in successive slices of a through-focus series, so that the last image taken in the series can have a significantly lower signal- to-noise ratio than the first. Bleaching is often successfully minimized by including antifade additives in the specimen preparation and/or by reducing the optical dose to the specimen. However, these measures may not be sufficient in a through-focus series where many slices must be taken. This paper presents a simple approach to ameliorating the effects of bleaching, which is to progressively increase the integration times so as to maintain constant signal level from slice to slice. I refer to the sequence of integration times as an integration schedule. I develop the equations for integration scheduling from the physical assumptions and discuss how the method affects image quality.

Paper Details

Date Published: 10 April 1997
PDF: 10 pages
Proc. SPIE 2984, Three-Dimensional Microscopy: Image Acquisition and Processing IV, (10 April 1997); doi: 10.1117/12.271273
Show Author Affiliations
Eric W. Hansen, Dartmouth College (United States)


Published in SPIE Proceedings Vol. 2984:
Three-Dimensional Microscopy: Image Acquisition and Processing IV
Carol J. Cogswell; Jose-Angel Conchello; Tony Wilson, Editor(s)

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