title: Single-Cell Isolation and Gene Analysis: Pitfalls and Possibilities.
abstract: During the last two decades single-cell analysis (SCA) has revealed extensive phenotypic differences within homogenous cell populations. These phenotypic differences are reflected in the stochastic nature of gene regulation, which is often masked by qualitatively and quantitatively averaging in whole tissue analyses. The ability to isolate transcripts and investigate how genes are regulated at the single cell level requires highly sensitive and refined methods. This paper reviews different strategies currently used for SCA, including harvesting, reverse transcription, and amplification of the RNA, followed by methods for transcript quantification. The review provides the historical background to SCA, discusses limitations, and current and future possibilities in this exciting field of research.
authors.affiliation: Department of Basic Sciences and Aquatic Medicine, Norwegian University of Life Sciences-Campus Adamstuen, 0033 Oslo, Norway. kjetil.hodne@nmbu.no.
authors.forename: Kjetil
authors.initials: K
authors.lastname: Hodne
authors.affiliation: Department of Basic Sciences and Aquatic Medicine, Norwegian University of Life Sciences-Campus Adamstuen, 0033 Oslo, Norway. finn-arne.weltzien@nmbu.no.
authors.forename: Finn-Arne
authors.initials: FA
authors.lastname: Weltzien
mesh_terms: D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D061346:Laser Capture Microdissection / Q000295:instrumentation / Q000379:methods; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods; D048348:Reverse Transcription; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D013824:Thermus / Q000737:chemistry
chemical_list: D012333:RNA, Messenger
keywords: Analysis; RT; amplification qPCR; harvesting; single-cell