Development of software and modification of Q-FISH protocol for estimation of individual telomere length in immunopathology

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Development of software and modification of Q-FISH protocol for estimation of individual telomere length in immunopathology. / Barkovskaya, M. S.; Bogomolov, A. G.; Knauer, N. Y. et al.

In: Journal of Bioinformatics and Computational Biology, Vol. 15, No. 2, 1650041, 01.04.2017.

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

BibTeX

@article{601a4fa5f78e487faf2db19b02fc6fbe,

title = "Development of software and modification of Q-FISH protocol for estimation of individual telomere length in immunopathology",

abstract = "Telomere length is an important indicator of proliferative cell history and potential. Decreasing telomere length in the cells of an immune system can indicate immune aging in immune-mediated and chronic inflammatory diseases. Quantitative fluorescent in situ hybridization (Q-FISH) of a labeled (C3TA2)3 peptide nucleic acid probe onto fixed metaphase cells followed by digital image microscopy allows the evaluation of telomere length in the arms of individual chromosomes. Computer-assisted analysis of microscopic images can provide quantitative information on the number of telomeric repeats in individual telomeres. We developed new software to estimate telomere length. The MeTeLen software contains new options that can be used to solve some Q-FISH and microscopy problems, including correction of irregular light effects and elimination of background fluorescence. The identification and description of chromosomes and chromosome regions are essential to the Q-FISH technique. To improve the quality of cytogenetic analysis after Q-FISH, we optimized the temperature and time of DNA-denaturation to get better DAPI-banding of metaphase chromosomes. MeTeLen was tested by comparing telomere length estimations for sister chromatids, background fluorescence estimations, and correction of nonuniform light effects. The application of the developed software for analysis of telomere length in patients with rheumatoid arthritis was demonstrated.",

keywords = "image analysis, immunopathology, integrated fluorescence intensity (IFI), peptide nucleic acid (PNA), Quantitative fluorescence in situ hybridization (Q-FISH), telomere length, RHEUMATOID-ARTHRITIS, HUMAN-CHROMOSOMES, IN-SITU HYBRIDIZATION, HUMANS, 17P, CANCER, DISEASES, FREQUENCY, SEQUENCE, Humans, Arthritis, Rheumatoid/genetics, In Situ Hybridization, Fluorescence/methods, Telomere, Telomere Homeostasis, Image Processing, Computer-Assisted/methods, Software",

author = "Barkovskaya, {M. S.} and Bogomolov, {A. G.} and Knauer, {N. Y.} and Rubtsov, {N. B.} and Kozlov, {V. A.}",

note = "Publisher Copyright: {\textcopyright} 2017 World Scientific Publishing Europe Ltd.",

year = "2017",

month = apr,

day = "1",

doi = "10.1142/S0219720016500414",

language = "English",

volume = "15",

journal = "Journal of Bioinformatics and Computational Biology",

issn = "0219-7200",

publisher = "World Scientific Publishing Co. Pte Ltd",

number = "2",

}

RIS

TY - JOUR

T1 - Development of software and modification of Q-FISH protocol for estimation of individual telomere length in immunopathology

AU - Barkovskaya, M. S.

AU - Bogomolov, A. G.

AU - Knauer, N. Y.

AU - Rubtsov, N. B.

AU - Kozlov, V. A.

PY - 2017/4/1

Y1 - 2017/4/1

N2 - Telomere length is an important indicator of proliferative cell history and potential. Decreasing telomere length in the cells of an immune system can indicate immune aging in immune-mediated and chronic inflammatory diseases. Quantitative fluorescent in situ hybridization (Q-FISH) of a labeled (C3TA2)3 peptide nucleic acid probe onto fixed metaphase cells followed by digital image microscopy allows the evaluation of telomere length in the arms of individual chromosomes. Computer-assisted analysis of microscopic images can provide quantitative information on the number of telomeric repeats in individual telomeres. We developed new software to estimate telomere length. The MeTeLen software contains new options that can be used to solve some Q-FISH and microscopy problems, including correction of irregular light effects and elimination of background fluorescence. The identification and description of chromosomes and chromosome regions are essential to the Q-FISH technique. To improve the quality of cytogenetic analysis after Q-FISH, we optimized the temperature and time of DNA-denaturation to get better DAPI-banding of metaphase chromosomes. MeTeLen was tested by comparing telomere length estimations for sister chromatids, background fluorescence estimations, and correction of nonuniform light effects. The application of the developed software for analysis of telomere length in patients with rheumatoid arthritis was demonstrated.

AB - Telomere length is an important indicator of proliferative cell history and potential. Decreasing telomere length in the cells of an immune system can indicate immune aging in immune-mediated and chronic inflammatory diseases. Quantitative fluorescent in situ hybridization (Q-FISH) of a labeled (C3TA2)3 peptide nucleic acid probe onto fixed metaphase cells followed by digital image microscopy allows the evaluation of telomere length in the arms of individual chromosomes. Computer-assisted analysis of microscopic images can provide quantitative information on the number of telomeric repeats in individual telomeres. We developed new software to estimate telomere length. The MeTeLen software contains new options that can be used to solve some Q-FISH and microscopy problems, including correction of irregular light effects and elimination of background fluorescence. The identification and description of chromosomes and chromosome regions are essential to the Q-FISH technique. To improve the quality of cytogenetic analysis after Q-FISH, we optimized the temperature and time of DNA-denaturation to get better DAPI-banding of metaphase chromosomes. MeTeLen was tested by comparing telomere length estimations for sister chromatids, background fluorescence estimations, and correction of nonuniform light effects. The application of the developed software for analysis of telomere length in patients with rheumatoid arthritis was demonstrated.

KW - image analysis

KW - immunopathology

KW - integrated fluorescence intensity (IFI)

KW - peptide nucleic acid (PNA)

KW - Quantitative fluorescence in situ hybridization (Q-FISH)

KW - telomere length

KW - RHEUMATOID-ARTHRITIS

KW - HUMAN-CHROMOSOMES

KW - IN-SITU HYBRIDIZATION

KW - HUMANS

KW - 17P

KW - CANCER

KW - DISEASES

KW - FREQUENCY

KW - SEQUENCE

KW - Humans

KW - Arthritis, Rheumatoid/genetics

KW - In Situ Hybridization, Fluorescence/methods

KW - Telomere

KW - Telomere Homeostasis

KW - Image Processing, Computer-Assisted/methods

KW - Software

UR - http://www.scopus.com/inward/record.url?scp=85010917537&partnerID=8YFLogxK

U2 - 10.1142/S0219720016500414

DO - 10.1142/S0219720016500414

M3 - Article

C2 - 28110603

AN - SCOPUS:85010917537

VL - 15

JO - Journal of Bioinformatics and Computational Biology

JF - Journal of Bioinformatics and Computational Biology

SN - 0219-7200

IS - 2

M1 - 1650041

ER -

ID: 8967226