ABSTRACT
Basic research in life science and medicine has dug into single cell level in recent years. Single-cell analysis offers to understand life from diverse perspectives and is used to profile cell heterogeneity to investigate mechanism of diseases. Single cell technologies have also found applications in forensic medicine and clinical reproductive medicine, while the techniques are rapidly evolving and have become more and more sophisticated. In this article, we reviewed various single cell isolation techniques and their pros and cons, including manual cell picking, laser capture microdissection and microfluidics, as well as analysis methods for DNA, RNA and protein in single cell. In addition, we summarized major up-to-date single cell research achievements and their potential applications.
Subject(s)
Animals , Cell Separation , DNA , Laser Capture Microdissection , RNA , Single-Cell AnalysisABSTRACT
Objective To develop a method to analyze functional grouping cDNA microarray gene expression in microvessels and neurons from rat brain using laser capture microdissection(LCM). Methods Microvessels and neurons were captured using the PixCell ⅡLCM instrument. The total RNA was extracted from the LCM samples according to the manufacturer's protocol of RNAqueous-Micro kit. The total RNA were processed for one round of T7-based RNA amplification; cDNA probe were synthesized using gene-specific primer and labeled for cDNA microarray analysis. Results Amplified RNA from microvessels and neurons allowed us to measure 96 gene expression in functional grouping cDNA microarray. Conclusion LCM allowed us to harvest pure microvessels and neurons from their native tissue environment, combined with methods of T7 RNA amplification and gene-specific primer amplification so that we can analyze functional grouping cDNA profiling in LCM samples.