Quantification and calibration of images in fluorescence microscopy

David S. Baskin; Marsha A. Widmayer; Martyn A. Sharpe

doi:10.1016/j.ab.2010.05.029

Quantification and calibration of images in fluorescence microscopy

David S. Baskin, Marsha A. Widmayer, Martyn A. Sharpe

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Fluorescence microscopy is a method widely used in life sciences to image biological processes in living and fixed cells or in fixed tissues. Quantification and calibration of images in fluorescence microscopy is notoriously difficult. We have developed a new methodology to prepare tissue " phantoms" that contain known amounts of (i) fluorophore, (ii) DNA, (iii) proteins, and (iv) DNA oligonucleotide standards. The basis of the phantoms is the ability of gelatin to act as a matrix for the conjugation of fluorophores as either a free-flowing liquid or a gelatinous solid depending on temperature (≥40 and ≤4°C).

Original language	English (US)
Pages (from-to)	118-126
Number of pages	9
Journal	Analytical Biochemistry
Volume	404
Issue number	2
DOIs	https://doi.org/10.1016/j.ab.2010.05.029
State	Published - Jan 1 2010

Keywords

Blunt-ended
Chemotherapy
Fluorescence microscopy
In situ
Ligation
Overhanging
Phantoms
Standards
U87

ASJC Scopus subject areas

Biochemistry
Biophysics
Molecular Biology
Cell Biology

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10.1016/j.ab.2010.05.029

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AU - Widmayer, Marsha A.

AU - Sharpe, Martyn A.

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AB - Fluorescence microscopy is a method widely used in life sciences to image biological processes in living and fixed cells or in fixed tissues. Quantification and calibration of images in fluorescence microscopy is notoriously difficult. We have developed a new methodology to prepare tissue " phantoms" that contain known amounts of (i) fluorophore, (ii) DNA, (iii) proteins, and (iv) DNA oligonucleotide standards. The basis of the phantoms is the ability of gelatin to act as a matrix for the conjugation of fluorophores as either a free-flowing liquid or a gelatinous solid depending on temperature (≥40 and ≤4°C).

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KW - Chemotherapy

KW - Fluorescence microscopy

KW - In situ

KW - Ligation

KW - Overhanging

KW - Phantoms

KW - Standards

KW - U87

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

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Quantification and calibration of images in fluorescence microscopy

Abstract

Keywords

ASJC Scopus subject areas

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