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PMID: 1614836 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Intracellular ion imaging using fluorescent dyes: artefacts and limits to resolution.

Pflugers Archiv : European journal of physiology ·Vol. 420 ·No. 5-6 ·1992-04-00 ·Pages 595-602

Silver RA, Whitaker M, Bolsover SR

Abstract

Development of highly efficient fluorescent ratio indicators has made imaging of ion concentrations within individual cells possible (Grynkiewicz et al. 1985; Tsien and Poenie 1986). Ion imaging is a complex technique and is therefore prone to artefacts. In this paper we investigate the limits of the technique and its potential pitfalls. The spatial resolution of an imaging system is determined for different cell geometries. We describe a technique to increase the time resolution of existing systems by using a single excitation wavelength to measure changes in ion concentration. We demonstrate examples of potential artefacts arising from hardware limitations, image processing and fundamental optics. Methods for recognition and minimization of these problems are discussed.

MeSH Terms
Calcium/metabolism Cytological Techniques Fluorescent Dyes Intracellular Fluid Microscopy, Fluorescence/methods
Chemicals
Fluorescent Dyes Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Silver R A
Department of Physiology, University College London, UK.
Whitaker M
Bolsover S R
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Article Info
Journal
Pflugers Archiv : European journal of physiology
Abbr.
Pflugers Arch
ISSN
0031-6768
DOI
10.1007/BF00374639
Published
1992-04-00
Pages
595-602
Language
English
Region
Germany
NLM ID
0154720
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source DOI: 10.1007/BF00374639 View journal details
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