Gene expression analysis in liver tissue at a single cell level by nested polymerase chain reaction and laser microdissection / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the measurements of gene expressing at a single hepatocyte level.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Combining laser microdissection and nested RT-PCR can monitor gene expression at a single cell level in vivo.</p>

Laser microdissection of a single cell from colon tissue for gene analysis / 中国应用生理学杂志

<p><b>AIM</b>To investigate the method of detecting gene expression in colon tissue at a single cell level.</p><p><b>METHODS</b>Individual cell(s) were picked up from colon frozen section using laser microdissection. RNA was extracted, reverse transcribed to complementary DNA (cDNA). cDNA was then analyzed by nested reverse transcription polymerase chain reaction (nested RT-PCR) using two pairs of primers.</p><p><b>RESULTS</b>Single cell(s) were selectively picked up using an ultraviolet laser micromanipulator. RNA was extracted, reverse transcribed and used for nested RT-PCR. Amplification products of cDNA from down to a single cell could be clearly visualized in the agarose gel.</p><p><b>CONCLUSION</b>The combined utilization of laser microdissection and nested RT-PCR provides an opportunity to analyze gene expression at single cell(s) level in colon tissue.</p>