Challenges of single-cell diagnostics: analysis of gene expression.

Analysis of single-cell gene expression promises a more precise understanding of human disease pathogenesis and important diagnostic applications. Here, we review the rationale for the study of gene expression at the single-cell level, practical methods to isolate homogeneous or single-cell samples, and current approaches to the analysis of single-cell gene expression. Finally, we highlight applications of laser microdissection-based gene expression analysis to the study of human disease and clinical diagnosis.

[1]  H. Höfler,et al.  Quantitative gene expression analysis in microdissected archival formalin-fixed and paraffin-embedded tumor tissue. , 2001, The American journal of pathology.

[2]  Gallya Gannot,et al.  Evaluation of non-formalin tissue fixation for molecular profiling studies. , 2002, The American journal of pathology.

[3]  D T Wong,et al.  Laser capture microdissection-generated target sample for high-density oligonucleotide array hybridization. , 2000, BioTechniques.

[4]  L. Liotta,et al.  Laser-capture microdissection: opening the microscopic frontier to molecular analysis. , 1998, Trends in genetics : TIG.

[5]  Michael G. Ormerod,et al.  Flow Cytometry: A Practical Approach , 1994 .

[6]  J. L. Stanton,et al.  Molecular phenotype of the human oocyte by PCR-SAGE. , 2000, Genomics.

[7]  E. Brown,et al.  Quantitative analysis of mRNA amplification by in vitro transcription. , 2001, Nucleic acids research.

[8]  A. Bashein,et al.  Global cDNA Amplification Combined with Real-Time RT–PCR: Accurate Quantification of Multiple Human Potassium Channel Genes at the Single Cell Level , 2000, Yeast.

[9]  D. Rappolee,et al.  Novel method for studying mRNA phenotypes in single or small numbers of cells , 1989, Journal of cellular biochemistry.

[10]  R. Bohle,et al.  Expression of angiotensin AT(1) and AT(2) receptors in adult rat cardiomyocytes after myocardial infarction. A single-cell reverse transcriptase-polymerase chain reaction study. , 2000, The American journal of pathology.

[11]  T. Freeman,et al.  Analysis of gene expression in single cells. , 1999, Current opinion in biotechnology.

[12]  J. Eberwine,et al.  Single-cell molecular biology: implications for the diagnosis and treatment of neurological disease. , 1999, Archives of neurology.

[13]  K. Kinzler,et al.  Analysing uncharted transcriptomes with SAGE. , 2000, Trends in genetics : TIG.

[14]  I. Black,et al.  Rab3A Is Required for Brain-Derived Neurotrophic Factor-Induced Synaptic Plasticity: Transcriptional Analysis at the Population and Single-Cell Levels , 2001, The Journal of Neuroscience.

[15]  M. Jackson,et al.  Gene expression profiles of laser-captured adjacent neuronal subtypes , 1999, Nature Medicine.

[16]  N. Iscove,et al.  Representative in Vitro cDNA Amplification From Individual Hemopoietic Cells and Colonies , 1990 .

[17]  N. Carter,et al.  Expression profiling of single cells using 3 prime end amplification (TPEA) PCR. , 1998, Nucleic acids research.

[18]  M. Kremer,et al.  Laser Capture Microdissection: Methodical Aspects and Applications with Emphasis on Immuno-Laser Capture Microdissection , 2001, Pathobiology.

[19]  L. Liotta,et al.  Laser Capture Microdissection , 1996, Science.

[20]  J. Eberwine,et al.  Amplification of mRNA populations using aRNA generated from immobilized oligo(dT)-T7 primed cDNA. , 1996, BioTechniques.

[21]  K. Schütze,et al.  Laser Microdissection as a New Approach to Prefertilization Genetic Diagnosis , 2001, Pathobiology.

[22]  Rudolf M. Huber,et al.  Combined transcriptome and genome analysis of single micrometastatic cells , 2002, Nature Biotechnology.

[23]  N. Datson,et al.  MicroSAGE: a modified procedure for serial analysis of gene expression in limited amounts of tissue. , 1999, Nucleic acids research.

[24]  Georgia Lahr,et al.  Identification of expressed genes by laser-mediated manipulation of single cells , 1998, Nature Biotechnology.