Fluorescent in situ hybridization FISH

Nucleic acid hybridization Fluorescent in situ hybridization (FISH)... 

A variation on this method, called fluorescent in situ hybridization (FISH), uses fluorescent-labeled DNA and RNA probes for detection and visualization of single cells by microscopy or flow cytometry.7 80 The FISH technique is popular because of its sensitivity and speed of visualization fluorescent dyes can be used to produce probes with different colors for simultaneous detection of several organisms.76,81,82... 

Moter, A. Gobel, U. B. Fluorescence in situ hybridization (FISH) for direct visualization of microorganisms. J. Microbiol. Meth. 2000, 41, 85-112. 

Recently, the use of AR has extended into several other areas, yielding interesting information for cytology, fresh cell/tissue sections, and fluorescence IHC (fluorescence in situ hybridization [FISH]), in addition to adaptations of the method for extraction of nucleic acids and proteins from FFPE tissues for use with modern methods of molecular analysis. In this chapter, the emphasis is on expanded applications in diagnostic cytology, fresh frozen cell/... 

Bouin s solution is one of the traditional ways to harden cell pellet. Some cytologists believe it provides the best cellular details, especially nuclear features in cell blocks.28 The major steps are (1) After centrifugation, fix the cell pellet with Bouin s solution. (2) After 2h, discard the solution. (3) Remove the hardened cell pellet from the tube, wrap it with lens paper, and transfer it into a cassette for further processing. We have been using this method for many years. In our experience, most of the time, ICC results are consistent with IHC from the surgical specimen. The biggest drawback of this method is the toxicity of Bouin s fixative which creates biohazard and safety issues for the laboratory. We also found cell blocks gave poor fluorescence in situ hybridization (FISH) results after Bouin s fixation. 

The RNA molecules, ribosomal RNA (rRNA) and messenger RNA (mRNA) play key roles in the protein synthesis. The amount of RNA in individual cells or in a community may, therefore, be taken as an indicator of protein synthesis and, thus, microbial activity. The number of active cells can be detected by fluorescent in situ hybridization (FISH) (Amann et al. 1995). By this method, individual cells carrying high concentrations of rRNA, situated on ribosomes, are quantified by fluorescence microscopy. The amount of rRNA in a community can also be detected by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), where rRNA extracted from soil is detected by creating a DNA copy and separating by gel electrophoresis (Duineveld et al. 2001). 

Total internal reflection fluorescence (TIRF) microscopy, fluorescence in situ hybridization (FISH), fluorescence recovery after photobleaching (FRAP), fluorescence lifetime imaging microscopy (FLIM). 

Advances in the fluorescence in situ hybridization (FISH) technique with the ability to label DNA probes with as many as six different spectrally distinct... 

Mass RD, Sanders C, Charlene K, et al. The concordance between the clinical trials assay (CTA) and fluorescence in situ hybridization (FISH) in the Herceptin pivotal trials. ProcAm Soc Clin Oncol 2000 19 75 (abstr). 

American College of Medical Genetics, Prenatal interphase fluorescence in situ hybridization (FISH) policy statement. Am. J. Hum. Genet. 53(2), 526-527 (1993). 

Gene amplification of the BCR-ABL 1 kinase may be associated with development of IM resistance. The presence of multiple copies of the BCR-ABL 1 gene in interphase nuclei can be demonstrated by fluorescence in situ hybridization (FISH) (40,42). In one series, more than half of the patients with IM resistance had evidence of clonal evolution with the development of additional chromosome abnormalities. Paired cytogenetic analyses performed at the begiiming of IM therapy and at the time of resistance demonstrated this evolution. 

Finally, several PNA based methods for use in genetic diagnostics have been developed. Of particular interest are methods for fluorescence in situ hybridization (FISH), modulation of polymerase chain reaction (PCR) analyses, and detection of genetic material by sensors and mass spectrometry. 8 ... 

Friedrich AB, Merkert H, Fendert T, Hacker J, Proksch P, Hentschel U (1999) Microbial Diversity in the Marine Sponge Aplysina cavernicola (formerly Verongia cavernicola) Analyzed by Fluorescence in Situ Hybridization (FISH). Mar Biol 134 461... 

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