Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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. и др.
в: Journal of Bioinformatics and Computational Biology, Том 15, № 2, 1650041, 01.04.2017.Результаты исследований: Научные публикации в периодических изданиях › статья › Рецензирование
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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.
N1 - Publisher Copyright: © 2017 World Scientific Publishing Europe Ltd.
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