Book Review

PATTERSON, B. K. (Editor)

Techniques in Quantification and Localization of Gene Expression

Basel, Berlin, Boston: Birkhäuser Verlag AG, 2000168 pages, 25 figures, 18 photomicrographs, 22 tables; DM 208.00ISBN 0-8176-4034-7

The subject of this little booklet is to present a variety of methods for the quantification andlocalization of gene expression within cells and tissues focussing mainly on the topic of viralinfection of human cells. All of these techniques cross disciplines, requiring knowledge of pathology,histochemistry and molecular biology.The primary factor to be considered in localization of gene expression is sensitivity. Sensitivity islargely dependent upon copy number. Detection of low abundance targets usually requires some typeof signal amplification. Some of these sensitive techniques allow single cell quantification of thetarget copies in specific cell types to be carried out. Others are qualitative at the cell population levelallowing the determination of the number of cells containing a particular target. Several approacheswere described in this book such as in situ PCR as well as the FISNA strategy (fluorescence in situ5'-nuclease assay), which avoids many potential pitfalls of the other method. Another technology, thein situ branched DNA assay (bDNA assay), was demonstrated to determine levels of viral nucleicacids by using schematically illustrated principal stages. Precisely elaborated protocols and step-by-step explanations facilitate reworking. A lot of trouble shooting was accomplished and a bunch oftricks were presented.Besides such techniques, it is mainly a book about basics like cell treatment, fixation, reducing ofautofluorescence and background staining as well as the selection of fluorescent dyes. A lot oftraditional methods were discussed, often forgotten and overlooked in their value for adequate samplepreparation. Certain reagent’s contribution to the staining of intracellular targets was clarified. Afurther main part of the booklet was stressed to technical flow cytometric details such as choice offilters and light sources as well as kinds of gating and compensation.Summarizing, it is a guide for experienced gene technologists who want to connect their procedureswith the cellular level, using flow cytometry and image analysis. Information is received about thecapacity of such methods to diagnose and track diseases like HIV and Hepatitis C.

S. MÜLLER

Acta Biotechnol. 21 (2001) 4, 332