In situ hybridization detection

Qian X, Bauer RA, Xu HS et al. In situ hybridization detection of calcitonin mRNA in routinely fixed, paraffin-embedded tissue sections a comparison of different types of probes combined with tyramide signal amplification. Appl Immunohistochem MolMorphol 2001 9 61-69. 

Table 2 summarizes transmitters/peptides observed in tufted cells. As noted, large populations of tufted cells contain the neuropeptide cholecystokinin (CCK) (Seroogy et al. 1985). One study using in situ hybridization detected substance P transcripts in some external tufted cells and in up to half of the mitral cells in MOB of rat (Warden... 

Bruch LA, Hill DA, Cai DX, et al. A role for fluorescence in situ hybridization detection of chromosome 22q dosage in distinguishing atypical teratoid/rhabdoid tumors from medulloblas-toma/central primitive neuroectodermal tumors. Hum Pathol. 2001 32 156-162. 

Enzyme-labeled fluorescence (ELF) Various phosphate-labeled fluorescent dyes, ELF 97 Uses phosphatase-based signal amplification. Applicable to immunohisto-chemical and cytological staining, mRNA in situ hybridization, detection of endogenous phosphatase activity, and blot analyses. 34... 

Deerinck, T. J., Martone, M. E., Lev-Ram, V., Green, D. P., Tsien, R. Y., Spector, D. L., Huang, S., and Ellisman, M. H. (1994). Fluorescence photooxidation with eosin A method for high resolution immunolocalization and in situ hybridization detection for light and electron microscopy. J. Cell Biol. 126, 901-910. 

A diagnostic method using fluorescence labeled DNA probes to detect and quantify the number complementary chromosomal sequences on a cellular resolution. A related technique that also allows assessment of gene amplifications, but without precise quantification of copy numbers is the chromogenic in situ hybridization (CISH). Here, instead of a fluorescent dye an enzyme that can generate a colored precipitate in the tissue samples is coupled to the DNA probe. 

PNA oligomers have also found widespread application as probes for in situ hybridization (FISH) both in human diagnostics [119-122] and in environmental detection of microbes [123, 124]. 

Perry-O Keeee H., Stender H., Broo-MER A., Oliveira K., Coull J., Hyldig-Nielsen J.J. Filter-based PNA in situ hybridization for rapid detection, identification and enumeration of specific microorganisms. /. Appl. Microbiol. 2001 90 180-189. 

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. 

With regard to the potential function of Raf-1 in mammalian germ cells, the testis is a site of abundant expression of c-ra/-l mRNAs. Northern blot and in situ hybridization analysis has shown that although c-ra/-l is expressed most abundantly in early pachytene spermatocytes, some transcripts were also detected in germ cells from type A and B spermatogonia through to the round spermatid stage (Wolfes et al., 1989 Wadewitz et al., 1993). 

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... 

Mittelman, M. W. Habash, M. Lacroix, J. M. Khoury, A. E. Krajden, M. Rapid detection of Enterobacteriaceae in urine by fluorescent 16S rRNA in situ hybridization on membrane filters. J. Microbiol. Meth. 1997, 30,153-160. 

Dirks RW, van Gijlswijk RP, Tullis RH et al (1990) Simultaneous detection of different mRNA sequences coding for neuropeptide hormones by double in situ hybridization using FITC- and biotin-labeled oligonucleotides. J Flistochem Cytochem 38 467 173... 

In situ hybridization Use of a DNA or RNA probe to detect the presence of the complementary DNA sequence in cloned bacterial or eukaryotic cells. 

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. 

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. 

Fluorescently labeled DNA probes can be used for detection, localization, or quantification of target DNA sequences. In situ hybridization mapping of genomic DNA sequences can be... 

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. 

Childs, C.V., Lloyd, J.M., Unabia, C., Gharib, S.D., Wierman, M.E., and Chin, W.W. (1987) Detection of luteinizing hormone b messenger ribonucleic acid (RNA) in individual gonadotropes after castration Use of a new in situ hybridization method with a photobiotinylated complementary RNA probe. Mol. Endocrinol. 1, 926-932. 

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. 

Antibody-based detection methods include immuno-cytochemistry, which gives qualitative data but has very good spatial resolution. Radioimmunoassays provide a quantitative measure of release or content. One of the major limitations of all antibody-based methods is the potential for cross-reactivity among the many peptides. For example, some of the most sensitive gastrin antisera also detect CCK, since the peptides share a common COOH-terminal tetrapeptide sequence. Methods for detection of the mRNAs encoding neuropeptides include Northern blots, which provide quantitative data and information on splice variants, but lack fine anatomical resolution. The more commonly used polymerase chain reaction, which can be quantitative but often is used in a more qualitative manner, provides great sensitivity. Alternatively, in situ hybridization preserves anatomical relationships and can be used to obtain both qualitative and quantitative data. 

Not detected High levels Dual-label in situ hybridization Hrabovszky et al. 2000... 

As an example of the use of antibodies labeled with alkaline phosphatase for detection of in situ hybridization, an infection with BNYVV virus in sugar beet is shown in Fig. 3C. Lectins labeled with an avidin-biotin fluorescein conjugate was used to visualize a-galactosyl groups on the surface of S. pombe in Fig. 3D. 

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