Fluorescence in-situ hybridization

In situ hybiidizatitMk has been detected using radioactive tracers and autoradiogn Ay. A moreomnirfetedesaiption can be found in many texts (see, e.g.. Ref. 63). Rn fluo rescmoe in situ hybridizatiMi, the probe DNA is made as 

The power of FISH is illustrated by its ability to identify all 24 human cfaiMnosomes. It is posaUe to devdop lAlA probes winch impart a unique ccdor to eadi of foe human duomoscNiies. Chromosome-painting jvobes are devdopedby FCR amplification DNA fragments from 

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Hongmanee P., Stender H., Rasmussen O.F. Evaluation of a fluorescence in situ hybridization assay for differentiation between tuberculous and nontuberculous Mycobacterium species in smears of Low-enstein-Jensen and mycobacteria growth indicator tube cultures using peptide nucleic acid probes. J. Clin. Microbiol. 2001 39 1032-1035. 

Drobniewski E.A., More P.G., Harris G. S. Differentiation of Mycobacterium tuberculosis complex and nontuberculous mycobacterial liquid cultures by using peptide nucleic acid-fluorescence in situ hybridization probes./. Clin. Microbiol. 2000 38 444-447. 

Fluorescence in situ hybridization Permits localization of a gene to one chromosomal band. 

FISH. Fluorescent in-situ hybridization a method utilizing fluorescently labeled DNA probes to detect or confirm gene or chromosome abnormalities that are generally beyond the resolution of routine cytogenetics. 

F15. Furuta, H., Nishi, S., Le Beau, M. M., Femald, A. A., Yano, H., and Bell, G. I Sequence of human hexokinase HI cDNA and assignment of the human hexokinase m gene (HK3) to chromosome band 5q35.2 by fluorescence in situ hybridization. Genomics 36,206-209 (1996). 

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/... 

Xia X, Rasmussen T, Alvarez X, et al. Fluorescence in situ hybridization using an old world monkey Y chromosome-specific probe combined with immunofluorescence staining on rhesus monkey tissues./. Histochem. Cytochem. 2007 55 1115-1121. 

Sugimura H. Detection of chromosome changes in pathology archives an application of microwave-assisted fluorescence in situ hybridization to human carcinogenesis studies. Carcinogenesis 2008 29 681-687. 

Watters AD, Bartlett MS. Fluorescence in situ hybridization in paraffin tissue sections. Mol. Biotechnol. 2002 21 217-220. 

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. 

Shin SJ, Chen B, Hyjek E, et al. Immunocytochemistry and fluorescence in situ hybridization in Her-2/neu status in cell block preparations. Acta Cytol. 2007 51 552-557. 

Beatty BG, Bryant R, Wang W, et al. Her-2/neu detection in fine-needle aspirates of breast cancer fluorescence in situ hybridization and immunocytochemical analysis. Am. J. Clin. Pathol. 2004 122 246-255. 

Bakkus, M.H., Brakel-van Peer, K.M., Adriaansem, H.J., Wierenga-Wolf, A.F., van den Akker, T.W., Dicke-Evingep M.J., and Benner, R. (1989) Detection of oncogene expression by fluorescent in situ hybridization in combination with immunofluorescent staining of cell surface markers. Oncogene 4,1255-1262. 

Meyne, J. (1993) Chromosome mapping by fluorescent in situ hybridization. In Methods in Nonradioactive Detection (G.C. Howard, ed.), pp. 263-268. Appleton Lange, Norwalk, Connecticut. 

No test has demonstrated reliable accuracy in the clinical setting for diagnosis of disseminated Candida infection. Blood cultures are positive in only 25% to 45% of neutropenic patients with disseminated candidiasis. Fluorescence in situ hybridization has excellent sensitivity and specificity in the identification of C. albicans from blood. 

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