Microfluidic extraction and stretching of chromosomal DNA from single cell nuclei for DNA fluorescence in situ hybridization.
We have developed a novel method for genetic characterization of single cells by integrating microfluidic stretching of chromosomal DNA and fiber fluorescence in situ hybridization (FISH). In this method, individually isolated cell nuclei were immobilized in a microchannel. Chromosomal DNA was released from the nuclei and stretched by a pressure-driven flow. We analyzed and optimized flow conditions to generate a millimeter-long band of stretched DNA from each nucleus. Telomere fiber FISH was successfully performed on the stretched chromosomal DNA. Individual telomere fiber FISH signals from single cells could be resolved and their lengths measured, demonstrating the ability of the method to quantify genetic features at the level of single cells.
Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, 2525 Pottsdamer Street, Tallahassee, FL 32310, USA.
Xiaozhu
X
Wang
Shin-Ichiro
S
Takebayashi
Evans
E
Bernardin
David M
DM
Gilbert
Ravindran
R
Chella
Jingjiao
J
Guan
D000818:Animals; D002467:Cell Nucleus / Q000235:genetics; D033481:Chromosomes, Mammalian; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D053595:Embryonic Stem Cells / Q000166:cytology; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods*; D016615:Telomere / Q000235:genetics / Q000378:metabolism
D004247:DNA