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Signal Amplification of FISH for Automated Detection Using Image Cytometry

The purpose of this study was to improve the detection of FISH signals, in order that spot counting by a fully automated image cytometer be comparable to that obtained visually under the microscope. Two systems of spot scoring, visual and automated counting, were investigated in parallel on stimulat...

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Detalles Bibliográficos
Autores principales: Truong, Khuong, Boenders, Jack, Maciorowski, Zofia, Vielh, Philippe, Dutrillaux, Bernard, Malfoy, Bernard, Bourgeois, Claire A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: IOS Press 1997
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4611095/
https://www.ncbi.nlm.nih.gov/pubmed/9223756
http://dx.doi.org/10.1155/1997/297216
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author Truong, Khuong
Boenders, Jack
Maciorowski, Zofia
Vielh, Philippe
Dutrillaux, Bernard
Malfoy, Bernard
Bourgeois, Claire A.
author_facet Truong, Khuong
Boenders, Jack
Maciorowski, Zofia
Vielh, Philippe
Dutrillaux, Bernard
Malfoy, Bernard
Bourgeois, Claire A.
author_sort Truong, Khuong
collection PubMed
description The purpose of this study was to improve the detection of FISH signals, in order that spot counting by a fully automated image cytometer be comparable to that obtained visually under the microscope. Two systems of spot scoring, visual and automated counting, were investigated in parallel on stimulated human lymphocytes with FISH using a biotinylated centromeric probe for chromosome 3. Signal characteristics were first analyzed on images recorded with a coupled charge device (CCD) camera. Number of spots per nucleus were scored visually on these recorded images versus automatically with a DISCOVERY image analyzer. Several fluochromes, amplification systems and pretreatments were tested. Our results for both visual and automated scoring show that the tyramide amplification system (TSA) gives the best amplification of signal if pepsin treatment is applied prior to FISH. Accuracy of the automated scoring, however, remained low (58% of nuclei containing two spots) compared to the visual scoring because of the high intranuclear variation between FISH spots.
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spelling pubmed-46110952016-01-12 Signal Amplification of FISH for Automated Detection Using Image Cytometry Truong, Khuong Boenders, Jack Maciorowski, Zofia Vielh, Philippe Dutrillaux, Bernard Malfoy, Bernard Bourgeois, Claire A. Anal Cell Pathol Other The purpose of this study was to improve the detection of FISH signals, in order that spot counting by a fully automated image cytometer be comparable to that obtained visually under the microscope. Two systems of spot scoring, visual and automated counting, were investigated in parallel on stimulated human lymphocytes with FISH using a biotinylated centromeric probe for chromosome 3. Signal characteristics were first analyzed on images recorded with a coupled charge device (CCD) camera. Number of spots per nucleus were scored visually on these recorded images versus automatically with a DISCOVERY image analyzer. Several fluochromes, amplification systems and pretreatments were tested. Our results for both visual and automated scoring show that the tyramide amplification system (TSA) gives the best amplification of signal if pepsin treatment is applied prior to FISH. Accuracy of the automated scoring, however, remained low (58% of nuclei containing two spots) compared to the visual scoring because of the high intranuclear variation between FISH spots. IOS Press 1997 1997-01-01 /pmc/articles/PMC4611095/ /pubmed/9223756 http://dx.doi.org/10.1155/1997/297216 Text en Copyright © 1997 Hindawi Publishing Corporation.
spellingShingle Other
Truong, Khuong
Boenders, Jack
Maciorowski, Zofia
Vielh, Philippe
Dutrillaux, Bernard
Malfoy, Bernard
Bourgeois, Claire A.
Signal Amplification of FISH for Automated Detection Using Image Cytometry
title Signal Amplification of FISH for Automated Detection Using Image Cytometry
title_full Signal Amplification of FISH for Automated Detection Using Image Cytometry
title_fullStr Signal Amplification of FISH for Automated Detection Using Image Cytometry
title_full_unstemmed Signal Amplification of FISH for Automated Detection Using Image Cytometry
title_short Signal Amplification of FISH for Automated Detection Using Image Cytometry
title_sort signal amplification of fish for automated detection using image cytometry
topic Other
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4611095/
https://www.ncbi.nlm.nih.gov/pubmed/9223756
http://dx.doi.org/10.1155/1997/297216
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