The peroxidase-mediated deposition of hapten- and fluorochrome-labeled tyramides has recently been shown to increase the sensitivity of immunofluorescence and fluorescence in situ hybridization techniques. We have evaluated a number of red, green, and blue fluorescent tyramides for detection of antigens in tissue sections and cytospin preparations and for the detection of hapten- and horseradish peroxidase-labeled probes hybridized in situ to cells and chromosomes. With few exceptions, all fluorescent tyramide-based methods provided a considerable increase in sensitivity compared to conventional immunofluorescence and FISH methods.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1177/002215549704500305 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Commentary on a Classic JHC Article on the Development of Highly Sensitive Fluorochrome-labeled Tyramides for Immunocytochemistry.
J Histochem Cytochem
May 2023
Department of Pathology and Cell Biology, Vagelos College of Physicians and Surgeons, Columbia University, New York, NY.
This commentary reflects on the significance and impact of the highly cited companion article that was published in the in 1997 (Gijlswijk RPM et al. Fluorochrome-labeled Tyramides: Use in Immunocytochemistry and Fluorescence In Situ Hybridization. .
View Article and Find Full Text PDFUse of fluorochrome-labeled rRNA targeted oligonucleotide probe and tyramide signal amplification to improve sensitivity of fluorescence in situ hybridization.
J Biosci Bioeng
October 2005
School of Bioresources, King Mongkut University of Technology Thonburi, Bangkhuntien, Bangkok 10150, Thailand.
A tyramide signal amplification (TSA) system was used in combination with a conventional fluorochrome-labeled 16S rRNA oligonucleotide probe to increase the sensitivity of fluorescence in situ hybridization. TSA was performed after hybridization resulted in a low fluorescence signal intensity. In contrast to the horseradish peroxidase-tyramide signal amplification (HRP-TSA) system and biotin-tyramide signal amplification (biotin-TSA) system, no additional expensive probe labeling was required.
View Article and Find Full Text PDFSimultaneous fluorescence in situ hybridization of mRNA and rRNA in environmental bacteria.
Appl Environ Microbiol
September 2004
Max-Planck-Institut für Marine Mikrobiologie, Celsiusstrabetae 1, D-28359 Bremen, Germany.
We developed for Bacteria in environmental samples a sensitive and reliable mRNA fluorescence in situ hybridization (FISH) protocol that allows for simultaneous cell identification by rRNA FISH. Samples were carbethoxylated with diethylpyrocarbonate to inactivate intracellular RNases and pretreated with lysozyme and/or proteinase K at different concentrations. Optimizing the permeabilization of each type of sample proved to be a critical step in avoiding false-negative or false-positive results.
View Article and Find Full Text PDFRapid synthesis of biotin-, digoxigenin-, trinitrophenyl-, and fluorochrome-labeled tyramides and their application for In situ hybridization using CARD amplification.
J Histochem Cytochem
June 1998
Department of Molecular Cell Biology and Genetics, University Maastricht, Maastricht, The Netherlands.
A one-step procedure for the synthesis of different tyramide conjugates, which can be utilized in the catalyzed reporter deposition (CARD) amplification system, is described. Succinimidyl esters of biotin, digoxigenin, and of the fluorochromes fluorescein, rhodamine, aminomethylcoumarine acetic acid, and Cy3 were coupled to tyramine in dimethylformamide (DMF) adjusted to a pH of 7.0-8.
View Article and Find Full Text PDFAn in situ hybridization histochemistry technique allowing simultaneous visualization by the use of confocal microscopy of three cellular mRNA species in individual neurons.
J Histochem Cytochem
June 1998
Laboratoire de Neuroendocrinologie Expérimentale, INSERM U 297 and Interactions Cellulaires Neuroendocriniennes, UMR 6544 CNRS, Institut Jean Roche, UER de Médecine Nord, Marseille, France.
We present a specific and sensitive method for simultaneous detection of three mRNA species in individual neurons. The method relies on the use of riboprobes labeled with [35S]-UTP, digoxigenin-UTP, or biotin-UTP. The nonradioactive probes were sequentially revealed by incubation with anti-digoxigenin immunoglobulins or streptavidin conjugated to peroxidase, followed by the use of fluorochrome-labeled tyramides as peroxidase substrates.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!