High-Throughput Human Telomere Length Analysis at the Single-Chromosome Level by FISH Coupled with Nano-Flow Cytometry

被引:3
作者
Gao, Kaimin [1 ]
Zhou, Yingxing [1 ]
Lu, Quanyi [2 ]
Lu, Jingyuan [2 ]
Su, Liyun [1 ]
Su, Rui [3 ]
Zhang, Miaomiao [1 ]
Tian, Ye [1 ]
Wu, Lina [1 ]
Yan, Xiaomei [1 ]
机构
[1] Xiamen Univ, Coll Chem & Chem Engn, Collaborat Innovat Ctr Chem Energy Mat, Dept Chem Biol,Key Lab Chem Biol Fujian Prov,MOE, Xiamen 361005, Peoples R China
[2] Xiamen Univ, Affiliated Zhongshan Hosp, Dept Hematol, Xiamen 361004, Peoples R China
[3] Xiamen Univ, Affiliated Hosp 1, Dept Hematol, Xiamen 361003, Peoples R China
基金
中国国家自然科学基金;
关键词
IN-SITU HYBRIDIZATION; METAPHASE CHROMOSOMES; FLUORESCENCE; QUANTIFICATION; NUMBER; DYNAMICS; CELLS; LOAD; SIZE; TIME;
D O I
10.1021/acs.analchem.1c01544
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Telomere length (TL) is a highly relevant biomarker for age-associated diseases and cancer, yet its clinical applications have been hindered by the inability of existing methods to rapidly measure the TL distribution and the percentage of chromosomes with critically short telomeres (CSTs, < 3 kb). Herein, we report the development of a high-throughput method to measure TL at the single-chromosome level. Metaphase chromosomes are isolated, hybridized with the Alexa Fluor 488-labeled telomeric peptide nucleic acid probe, and analyzed using a laboratory-built ultrasensitive nano-flow cytometer. The fluorescence intensity of individual chromosomes is converted to TL in kilobases upon external calibration. With an analysis rate of several thousand chromosomes per minute, a statistically robust TL distribution histogram is acquired in minutes, and the percentage of chromosomes with CSTs can be quickly assessed. By analyzing peripheral blood lymphocytes of 158 healthy donors, TL is found to shorten with age at a rate of 64 +/- 3 bp/year and the percentage of chromosomes with CSTs increases with age at a rate of 0.32 +/- 0.02%/year. Moreover, the data of 28 patients with chronic myeloid leukemia (CML) indicate that telomeres are significantly shorter at the time of diagnosis and the clinical phases of CML are closely associated with TL and the percentage of chromosomes with CSTs. This powerful tool could greatly deepen our understanding of telomere biology and improve the clinical utility of telomere biomarkers.
引用
收藏
页码:9531 / 9540
页数:10
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