[An application of laser capture microdissection to isolate the pure atrial cells].
OBJECTIVE
To use the laser capture microdissection (LCM) to isolate the pure atrial cells from heart. To extract tiny amount RNA and amplify the genes of beta-actin and GAPDH.


METHODS
First, the prepared sample RNA was extracted and verified. Second, the frozen sections were scraped after haematoxylin-eosin staining, and then the RNA of scraped tissues was extracted and verified. Last, the tiny amount RNA was extracted from atrial cells captured by LCM, and the genes of beta-actin and GAPDH were amplified by reverse transcriptase polymerase chain reaction (RT-PCR).


RESULTS
The atrial cells isolated with LCM had clear morphology after quick staining. The high quality RNA was extracted from LCM sample and scraped tissues, of which A260/A280 was 1.9-2.0. And 18S rRNA and 28S rRNA were seen distinctly on the gel of RNA formaldehyde denaturing gel electrophoresis, which indicated that RNA didn't degrade before cells captured. Too small amount of RNA extracted from captured cells was below the detectable limit of gel electrophoresis or ultraviolet spectrophotometer. Bands of 300 bp and 357 bp could be amplified by RT-PCR from tiny amount RNA.


CONCLUSIONS
Highly pure atrial cells could be obtained by LCM. Tiny amount RNA of captured atrial cells could be extracted and amplified with genes of beta-actin and GAPDH.
Ou
Yan
Y
Department of Cardiology, Second Hospital of Xi'an Jiaotong University, Xi'an 710004, China.
Niu
Xiao-lin
XL
Huang
Chen
C
Ren
Fu-xian
FX
Chen
Wei
W
D000199:Actins / Q000235:genetics; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D006325:Heart Atria / Q000166:cytology*; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D012313:RNA / Q000302:isolation & purification; D051381:Rats
D000199:Actins; D012313:RNA