Background Information

Combined immunophenotyping and FISH

Combined genotype/phenotype analysis provides information about the genetic composition and identity of cells present in a heterogeneous population. This type of information has diagnostic and prognostic value for tumor cytogeneticists and hematologists studying blood cancers. Additional information concerning the expression levels of genes of interest can be obtained by combining an RNA-FISH procedure with immunophenotyping. These techniques can be applied to cell preparations, and in principle also to tissue sections (Weber-Matthiesen et al., 1993). The difficulty with tissue sections is, however, the high level of autofluorescence that is often encountered. Furthermore, it should be noted that due to the large variety in immunophenotypic markers and cell types, it is impossible to provide a general protocol applicable to all of them. Therefore, in practice, different types of fixation and possibly preparation techniques should be tested. The number of reports describing combined immunophenotyping and and FISH is increasing, however, and valuable practical information can be found there for specific applications (e.g., Speel et al., 1994; Hessel et al., 1996; Litle et al., 1996).

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