Method for selectively staining chitin-containing organisms
AssigneeUniversity of South Florida
InventorMy Lien Dao
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Abstract
The present invention provides a method for the staining of fungi and microsporidia for observation with a light microscope based upon the presence of chitin in the composition of these organisms. With the method of the present invention a sample to be analyzed is treated with a solution of Ponceau S and Stains-all dye. The sample is then selectively decolorized and rinsed. The resulting sample is examined with a light microscope, or photographed for a permanent record, to identify the presence of a variety of microorganisms, to include fungi and microsporidia.
Claims10
A method for determining the presence or absence of a chitin-containing microorganism in a sample suspected of containing a chitin-containing microorganisms, the method comprising the steps of:
- providing a sample suspected of containing a chitin-containing microorganisms to be analyzed;
- treating the sample with 3-hydroxy-4((2-sulfo-4-((-4-sulfophenyl(azo)phenyl)azo) 2,7 naphthalenedisulfonic acid tetrasodium salt stain to stain the chitin of the chitin-containing microorganisms;
- treating the sample with a cationic carbocyanine 1-Ethyl-2-[3-(1-ethylnaphtho[1,2-d]thiazolin-2-ylidene)-2-methylpropenyl]naphto[1,2-d]thiazolium bromide dye, the cationic carbocyanine dye binding to the 3-hydroxy-4((2-sulfo-4-((-4-sulfophenyl(azo)phenyl)azo) 2,7 naphthalenedisulfonic acid tetrasodium salt stain to form a light-stable complex;
- applying three decolorizing solutions to the sample;
- rinsing the sample after each application of decolorizing solution;
- and
- examining the sample with a light microscope to identify the presence of chitin-containing microorganisms in the sample, wherein the stained chitin-containing microorganisms are observable in the light microscope.
A method for determining the presence or absence of chitin-containing fungi or microsporidia, the method comprising the steps of:
- providing a sample suspected of containing a chitin-containing fungi or microsporidia to be analyzed;
- treating the sample with 3-hydroxy-4((2-sulfo-4-((-4-sulfophenyl(azo)phenyl)azo) 2,7 naphthalenedisulfonic acid tetrasodium salt stain to stain the chitin of the chitin-containing fungi or microsporidia;
- treating the sample with 1-Ethyl-2-[3-(1-ethylnaphtho[1,2-d]thiazolin-2-ylidene)-2-methylpropenyl]naphto[1,2-d]thiazolium bromide dye, which dye binds to 3-hydroxy-4((2-sulfo-4-((-4-sulfophenyl(azo)phenyl)azo) 2,7 naphthalenedisulfonic acid tetrasodium salt stain to form a light-stable complex;
- applying three decolorizing solutions to the sample;
- rinsing the sample after each application of decolorizing solution;
- and
- examining the sample with a light microscope to identify the presence of chitin-containing fungi or microsporidia in the sample, wherein the stained chitin-containing fungi or microsporidia are observable in the light microscope.
Dependencies are inferred from the claim text.