Higher axial-resolution and sensitivity pachytene fluorescence in situ hybridization protocol in tetraploid cotton

2009 ◽  
Vol 17 (8) ◽  
pp. 1041-1050 ◽  
Author(s):  
Kai Wang ◽  
Zaijie Yang ◽  
Changshen Shu ◽  
Jing Hu ◽  
Qiuyun Lin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Tetraploid Cotton ◽  
Axial Resolution ◽  
Hybridization Protocol

scholarly journals Catalyzed Reported Deposition-Fluorescence In Situ Hybridization Protocol To Evaluate Phagotrophy in Mixotrophic Protists

2005 ◽  
Vol 71 (11) ◽  
pp. 7321-7326 ◽  
Author(s):  
Juan M. Medina-Sánchez ◽  
Marisol Felip ◽  
Emilio O. Casamayor
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Signal Amplification ◽  
Cell Attachment ◽  
Cell Loss ◽  
Alexa Fluor ◽  
Hybridization Protocol ◽  
Card Fish ◽  
First Time

ABSTRACT We describe a catalyzed reported deposition-fluorescence in situ hybridization (CARD-FISH) protocol particularly suited to assess the phagotrophy of mixotrophic protists on prokaryotes, since it maintains cell and plastid integrity, avoids cell loss and egestion of prey, and allows visualization of labeled prey against plastid autofluorescence. This protocol, which includes steps such as Lugol's-formaldehyde-thiosulfate fixation, agarose cell attachment, cell wall permeabilization with lysozyme plus achromopeptidase, and signal amplification with Alexa-Fluor 488, allowed us to detect almost 100% of planktonic prokaryotes (Bacteria and Archaea) and, for the first time, to show archaeal cells ingested by mixotrophic protists.

scholarly journals A modified fluorescence in situ hybridization protocol for Plasmodium falciparum greatly improves nuclear architecture conservation

2010 ◽  
Vol 173 (1) ◽  
pp. 48-52 ◽  
Author(s):  
Monica Contreras-Dominguez ◽  
Carolina Borsoi Moraes ◽  
Thierry Dorval ◽  
Auguste Genovesio ◽  
Fernando de Macedo Dossin ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Nuclear Architecture ◽  
Hybridization Protocol

Direct and sensitive fluorescence in situ hybridization of 45S rDNA on tomato chromosomes

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1062-1065 ◽  
Author(s):  
Jie Xu ◽  
E. D. Earle
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome 2 ◽  
45S Rdna ◽  
Plant Chromosomes ◽  
Signal Sensitivity ◽  
Direct Fluorescence ◽  
Hybridization Protocol ◽  
Tomato Genotypes

We describe a direct and sensitive fluorescence in situ hybridization protocol for plant chromosomes. We labelled 45S rDNA with fluorescein-12-dUTP and hybridized to somatic chromosomes of four tomato genotypes. This technique does not require posthybridization immunocytochemical amplifications. The improved signal sensitivity with this technique allowed identification of new rDNA loci on three pairs of chromosomes, in addition to the previously known locus on chromosome 2. We discuss favorable features of direct fluorescence in situ hybridization for chromosomes fixed on a slide and chromosomes or cells in suspension.Key words: direct fluorescence in situ hybridization, 45S rDNA, tomato chromosomes.

Development of a Fluorescence In Situ Hybridization protocol for analysis of Gram-positive and Gram-negative bacteria

2016 ◽  
Vol 33 (3) ◽  
pp. 434
Author(s):  
Marina González-Pérez ◽  
Rafaela Fernandes ◽  
Ricardo Vieira ◽  
António Pereira ◽  
António Candeias ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Gram Negative Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Hybridization Protocol

Colocalization of mRNAs by Fluorescence in Situ Hybridization

1999 ◽  
Vol 5 (S2) ◽  
pp. 480-481
Author(s):  
D. G. Baskin ◽  
J. F. Breininger
Keyword(s):  
In Situ Hybridization ◽  
Nucleic Acid ◽  
Fluorescence In Situ Hybridization ◽  
Radioactive Isotopes ◽  
Nucleic Acid Probes ◽  
Messenger Ribonucleic Acid ◽  
Hybridization Probes ◽  
Isotopic Methods ◽  
Hybridization Protocol

Fluorescence in situ hybridization (FISH), long the method of choice for chromosomal cytogenetics, is becoming recognized as a powerful method for correlative histochemical detection of multiple messenger ribonucleic acid (mRNA) in cells. The technique is based upon the principle of the binding of a labeled strand of DNA (an oligonucleotide probe) or RNA (a riboprobe) to complementary strands of mRNA. In the traditional in situ hybridization method, nucleic acid probes are labeled with radioactive isotopes and the hybrids are localized by autoradiography. More recently, labeling of the probes with digoxigenin, biotin, or fluorescein has allowed nonisotopic detection of probe-mRNA hybrids with immunoenzymatic techniques that are based on peroxidase or alkaline phosphatase histochemistry, in which the results are observed as brightfield stains. FISH is similar to conventional nonisotopic in situ hybridization except that the labeled nucleic acid probes are detected by fluorescence microscopy.The FISH technique has advantages for performing correlative histochemistry. The results of a FISH protocol can be observed within a few hours of completing the hybridization protocol instead of waiting for days as is the case with isotopic methods. FISH permits cellular resolution and facilitates multiple labeling, where in situ hybridization probes can be resolved at the level of individual cells (Hahn et al. 1998).

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