http://rdf.ncbi.nlm.nih.gov/pubchem/patent/GB-1319710-A

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filingDate 1971-07-12-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationDate 1973-06-06-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber GB-1319710-A
titleOfInvention Blood analysis method and composition
abstract 1319710 Dye composition for distinguishing blood cells BIO/PHYSICS SYSTEMS INC 12 July 1971 [13 July 1970] 32556/71 Heading B1X [Also in Divisions C4 G1 and G4] In the analysis of blood samples in order to distinguish white from other cells, the samples are combined with a fluorescent dye which is absorbed mainly by the white cells. The dye may be acridine orange (3, 6-bisdimethyl-amino-acridinium-choride) in aqueous solution in a concentration between 10<SP>-7</SP> and 10<SP>-5</SP> gms per cc., and having a pH factor and an osmolality within the normal physiological ranges for human blood plasma. The pH factor may be obtained by addition of phosphates e.g. NaH 2 PO 4 and Na 2 HPO 4 and the osmolality adjusted by addition of sodium chloride e.g. to give a 0.85% solution. A method of producing a solution such as the latter is described. Instead of the phosphate buffer, HEPES (N-2-hydroxyethylpiperazine- N'-2-ethanesulfonic acid), or TRIS (2-amino- 2-(hydroxy-methyl)-1, 3-propanediol), may be used. Examples of various concentrations of acridine orange solution were given. The dye is characterised by a green fluorescence for white cells and red for granulocytic cells. Other dyes are mentioned which produce a fluorescent effect with cells:-phosphine (very mild-yellow for white cells) and (even more mild fluorescence) phenosafranine, neutral red, auromine O and acradine yellow.
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