FISHing in the microwave: the easy way to preserve proteins. I. Colocalization of DNA probes and surface antigens in human leukocytes.
Most of the techniques for simultaneous visualization of proteic cell components and DNA probes are difficult to carry out. We have developed an alternative protocol for simultaneous visualization of DNA probes and cell surface antigens in human cells. The method exploits microwave energy to denature DNA and to anneal the probe after antigen fluorescent detection of unfixed cells previously embedded in a microgel. CD3 antigens and whole genome DNA probes or specific repetitive DNA sequences were colocalized in peripheral blood samples. The results show a strong, specific and consistent hybridization pattern in each cell that allowed correlation between cell subtype (PROTEINS) and nuclear phenotype (DNA).
Gosálvez
J
J
Departamento de Biologia, Universidad Autónoma de Madrid, Spain. jaime.gosalvez@uam.es
De La Torre
J
J
Pita
M
M
Martinez-Ramirez
A
A
López-Fernandez
C
C
Goyanes
V
V
López-Fernández
J
J
D017252:CD3 Complex / Q000378:metabolism*; D015342:DNA Probes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007962:Leukocytes / Q000378:metabolism*; D008872:Microwaves
D017252:CD3 Complex; D015342:DNA Probes