Feulgen And Mann's Reaction
   HOME

TheInfoList



OR:

Feulgen stain is a
staining Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology (microscopic study of biological tissues), in cytology (microscopic study of cells), and in the ...
technique discovered by
Robert Feulgen Joachim Wilhelm Robert Feulgen (2 September 1884 – 24 October 1955) was a German physician and chemist who, in 1914, developed a method for staining DNA (now known as the Feulgen stain) and who also discovered plant and animal nuclear Nuclear ...
and used in histology to identify chromosomal material or DNA in cell specimens. It is darkly stained. It depends on acid hydrolysis of DNA, therefore fixating agents using strong acids should be avoided. The specimen is subjected to warm (60 °C) hydrochloric acid, then to Schiff reagent. In the past, a
sulfite Sulfites or sulphites are compounds that contain the sulfite ion (or the sulfate(IV) ion, from its correct systematic name), . The sulfite ion is the conjugate base of bisulfite. Although its acid ( sulfurous acid) is elusive, its salts are wide ...
rinse followed, but this is now considered unnecessary. Optionally, the sample can be
counterstain A counterstain is a stain with colour contrasting to the principal stain, making the stained structure easily visible using a microscope. Examples include the malachite green counterstain to the fuchsine stain in the Gimenez staining technique a ...
ed with
Light Green SF yellowish Light green SF, also called C.I. 42095, light green SF yellowish, is a green triarylmethane dye. Uses Biomedical It is used in histology for staining collagen; for that purpose it is a standard dye in North America. In Masson's trichrome it i ...
. Finally, it is dehydrated with ethanol, cleared with xylene, and mounted in a resinous medium. DNA should be stained red. The background, if counterstained, is green. The Feulgen reaction is a semi-quantitative technique. If the only aldehydes remaining in the cell are those produced from the hydrolysis of DNA, then the technique is quantitative for DNA. It is possible to use an instrument known as a microdensitometer or microspectrophotometer to actually measure the intensity of the pink Feulgen reaction for a given
organelle In cell biology, an organelle is a specialized subunit, usually within a cell, that has a specific function. The name ''organelle'' comes from the idea that these structures are parts of cells, as organs are to the body, hence ''organelle,'' the ...
. Using this procedure, it was early determined that interphase cells were composed of two populations, those with
diploid Ploidy () is the number of complete sets of chromosomes in a cell, and hence the number of possible alleles for autosomal and pseudoautosomal genes. Sets of chromosomes refer to the number of maternal and paternal chromosome copies, respectively ...
DNA and those with tetraploid DNA (two complete genomes). The nuclei looked identical, but one contained twice as much DNA. This gave rise to the division of the interphase period of the cell cycle to G1, S, and G2 phases based on the synthesis of that extra DNA.


References

{{Reflist Biochemistry detection reactions Microscopy Microbiology techniques Laboratory techniques Staining Histotechnology Staining dyes