[Gene expression analysis in liver tissue at a single cell level by nested polymerase chain reaction and laser microdissection].
OBJECTIVE
To investigate the measurements of gene expressing at a single hepatocyte level.


METHODS
Individual hepatocyte was isolated from cryostat tissue section using laser microdissection technique. To detect the mRNA expressed by single hepatocyte, RNA was extracted, reversely transcribed to cDNA and amplified by nested polymerase chain reaction (PCR).


RESULTS
Single cell was microdissected from cryostat tissue using an ultraviolet laser micromanipulator. The RNA could be extracted from the isolated cell(s), and the RT-PCR production could be observed after electrophoresis, whose quantitation was compatible with the number of cells.


CONCLUSIONS
Combining laser microdissection and nested RT-PCR can monitor gene expression at a single cell level in vivo.
Shi
Xin
X
Department of General Surgery, Zhongda Hospital, Southeast University, Nanjing 210009, China.
Gao
Nai-rong
NR
Huo
Ming-dong
MD
Hu
Hao-lin
HL
Friess
Helmut
H
Buchler
Markus
M
D004210:Dissection; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D007834:Lasers; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012333:RNA, Messenger