In Situ Hybridization in Neurobiology: Advances in Methodology
James H. Eberwine, Karen L. Valentino, Jack D. Barchas
Oxford University Press, 1994 - Medical - 199 pages
In situ hybridization has become an important and widely used research tool for neurobiologists since the authors' first book on the subject, In Situ Hybridization: Applications to Neurobiology (Oxford, 1987). This new volume examines the methodological advances that have been made in in situ hybridization techniques and their impact on the study of the nervous system. These include methods for improving sensitivity, ways of examining multiple mRNAs within the same section, ways of examining both proteins and mRNAs within the same section, and methods for using in situ hybridization technology to examine areas of cellular biological significance, such as translational control. The development of alternative labels to radioactivity for the detection of hybridized probes, and the combined use of in situ hybridization and immunocytochemistry is also covered in this book. It discusses the quantification of in situ hybridization signal and presents an overview of information generated by using in situ hybridization in diagnostic medicine. This volume will be essential to researchers in the neurosciences as well as those in other disciplines.
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Methodological Considerations in the Utilization of In Situ
Combining Nonisotopic In Situ Hybridization Histochemistry with Steroid
Development of Techniques to Combine Isotopic and Nonisotopic In Situ
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2X SSC 30 minutes amplification analysis antibody aRNA assay autoradiography Baldino binding biotin biotinylated bridization cDNA cDNA synthesis cellular Chesselet clones colocalization concentration containing coverslips cRNA probes denatured DEPC DEPC-treated detection digoxigenin digoxigenin-labeled dNTPs dual-label ISHH Eberwine Emson emulsion encoding enzyme estrogen receptor ethanol exposure Figure fixed formamide GABAA receptor gene expression Haase histochemistry hormone hybridization histochemistry hybridization probes hybridization signal hypothalamus incubation individual cells infection IS-PCR Kiyama levels of labeling LHRH LHRH mRNA LHRH neurons method molecular mRNA mRNA levels NaCl neurons Neuroscience nonisotopic nonradioactive nonspecific Northern blots nucleic acid nucleotides oligonucleotide probes paraformaldehyde peptide pituitary polymerase chain reaction POMC prehybridization primary transcript primer procedure proopiomelanocortin protein quantitative radiolabeled receptor subunit region reverse transcriptase riboprobes RNA polymerase RNase room temperature sensitivity sequence silver grains slides sodium solution staining stem-loop steroid studies synthetic oligonucleotide technique tissue sections Toran-Allerand washed