Deconvolving single-molecule intensity distributions for quantitative microscopy measurements.

Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Research Abstract Details 

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Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Abstract Text:

Sarah A Mutch,Bryant S Fujimoto,Christopher L Kuyper,Jason S Kuo,Sandra M Bajjalieh,Daniel T Chiu,

In fluorescence microscopy, images often contain puncta in which the fluorescent molecules are spatially clustered. This article describes a method that uses single-molecule intensity distributions to deconvolve the number of fluorophores present in fluorescent puncta as a way to "count" protein number. This method requires a determination of the correct statistical relationship between the single-molecule and single-puncta intensity distributions. Once the correct relationship has been determined, basis histograms can be generated from the single-molecule intensity distribution to fit the puncta distribution. Simulated data were used to demonstrate procedures to determine this relationship, and to test the methodology. This method has the advantages of single-molecule measurements, providing both the mean and variation in molecules per puncta. This methodology has been tested with the avidin-biocytin binding system for which the best-fit distribution of biocytins in the sample puncta was in good agreement with a bulk determination of the avidin-biocytin binding ratio.

Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Publishing Authors By Initials

SA Mutch,BS Fujimoto,CL Kuyper,JS Kuo,SM Bajjalieh,DT Chiu,

For similar nervous system: synapses: synaptic vesicles research abstracts see: nervous system: synapses: synaptic vesicles research

PUBMED ID PMID:

MEDLINE DATE:

Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Biophysical journal

VOLUME: 92

Page Numbers: 2926-43

Journal Abbreviation: Biophys. J.

ISSN: 0006-3495

DAY: 26

MONTH: 01

YEAR: 2007

Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 370626

Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Keywords Mesh Terms:

KEYWORDS: Synaptic Vesicles

MESH TERMS: ultrastructure

Chemical & Substance for Abstract: Deconvolving single-molecule intensity distributions for quantitative microscopy measurements. Information

Substance Name: biocytin

Registry Number: 576-19-2

Grant and Affiliation Information for Deconvolving single-molecule intensity distributions for quantitative microscopy measurements.

AFFILIATION: Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.

Country: United States

AGENCY: United States NINDS

GRANT: NS052637

ACRONYM: NS

MEDLINETA: Biophys J

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