I prepared aceto-orecin stain in glacial acetic acid (later adding the HCl) and I am visualizing plant roots which are in Murashige Skoog Media. The protocol which I was interested in is from Dubrovsky 2006 which utilized Neutral Red. I only have orcein in my lab currently and I would like to know if I can prepare an Orcein stain in 0.2X MS media with Phosphate buffer similar to what is mentioned in the paper. The pH of the phoshpate buffer is 8, and MS media is 5.8, aceto-orcein is generally acidic as far as I know. I read that orcein is generally used for chromosome visualization in plant cells. But can it be used in liquid Murashige and Skoog media for visualization of root hair growth?
Hello,
please read this link :
https://en.wikipedia.org/wiki/Orcein
and give infos about your staining method (Plant roots in vivo /Water solution ...) for the answerers.
Orcein is not approved as a food dye (banned in Europe since January 1977), with E number E121 before 1977 and E182 after.[5],[6] Its CAS number is 1400-62-0. Its chemical formula is C28H24N2O7. It forms dark brown crystals. It is a mixture of phenoxazone derivates - hydroxyorceins, aminoorceins, and aminoorceinimines.
The chemical components of orcein were elucidated only in the 1950s by Hans Musso.[7] The structures are shown below. A paper originally published in 1961, embodying most of Musso's work on components of orcein and litmus, was translated into English and published in 2003[8] in a special issue of the journal Biotechnic & Histochemistry (Vol 78, No. 6) devoted to the dye. A single alternative structural formula for orcein, possibly incorrect, is given by the National Library of Medicine [1] and Emolecules [2].
Orcein is a reddish-brown dye, orchil is a purple-blue dye. Orcein is also used as a stain in microscopy to visualize chromosomes,[9] elastic fibers,[10] Hepatitis B surface antigens,[11] and copper-associated proteins.[12]
Solubility and pH for orcein staining are important for your future research.
JRG
Thank you for your suggestions. I read that orcein is generally used for chromosome visualization in plant cells. But can it be used in liquid Murashige and Skoog media for visualization of root hair growth?
What is the exact role of orcein while being used for chromosomal visualizations? Think on these lines, roots are organized structure, Where exactly orcein binds during chromosomal work , is there any chemical or physical reaction and can it work when used in MS medium to root hairs? think on these lines, if no answer, then why dont u play once.
According to the above references posted in my answer, Prof. Musso found and separated 14 dyes of orcein chromatographically.
If you buy the commercial orcein, you must do a TLC and then separate the compounds vis CC (column Chromatograpy).
Allah Bakhsh is right in his comments and his posted questions in his comments can help you to investigate your problem in a scientific manner.
( p.Ex. )
Where exactly orcein binds during chromosomal work , is there any chemical or physical reaction and can it work when used in MS medium to root hairs?
The answer to this question can be found in the Muss's paper if you consider the cited isomers and its pK values. Choose one of them and search in Gogle (e.g.
aminophenoxazone or hydroxyphenoxazon). Test them and compare the chromosomal staining.
I prefer INTERACTION for the binding and binding site.
My suggestion : try to synthesize the cited isomers in my answer and test them .
You will have ONE COMPOUND for staining and not a MIXTURE.
JRG
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