Flow cytometry and FISH to measure the average length of telomeres (flow FISH)

Baerlocher, Gabriela M; Vulto, Irma; de Jong, Gary; Lansdorp, Peter M (2006). Flow cytometry and FISH to measure the average length of telomeres (flow FISH). Nature physics, 1(5), pp. 2365-76. Basingstoke: Nature Publishing Group 10.1038/nprot.2006.263

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Telomeres have emerged as crucial cellular elements in aging and various diseases including cancer. To measure the average length of telomere repeats in cells, we describe our protocols that use fluorescent in situ hybridization (FISH) with labeled peptide nucleic acid (PNA) probes specific for telomere repeats in combination with fluorescence measurements by flow cytometry (flow FISH). Flow FISH analysis can be performed using commercially available flow cytometers, and has the unique advantage over other methods for measuring telomere length of providing multi-parameter information on the length of telomere repeats in thousands of individual cells. The accuracy and reproducibility of the measurements is augmented by the automation of most pipetting (aspiration and dispensing) steps, and by including an internal standard (control cells) with a known telomere length in every tube. The basic protocol for the analysis of nucleated blood cells from 22 different individuals takes about 12 h spread over 2-3 days.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Haematology, Oncology, Infectious Diseases, Laboratory Medicine and Hospital Pharmacy (DOLS) > Clinic of Haematology and Central Haematological Laboratory

UniBE Contributor:

Baerlocher, Gabriela M.

ISSN:

1745-2473

ISBN:

17406480

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Factscience Import

Date Deposited:

04 Oct 2013 14:50

Last Modified:

17 Mar 2015 21:53

Publisher DOI:

10.1038/nprot.2006.263

PubMed ID:

17406480

Web of Science ID:

000251155600025

URI:

https://boris.unibe.ch/id/eprint/21182 (FactScience: 5158)

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