US 12,072,330 B2
Method for classifying/counting leukocytes, reagent kit for classifying leukocytes, and reagent for classifying leukocytes
Saori Otsuka, Kusatsu (JP); Kazuki Hatcho, Kobe (JP); Toshihiro Mizukami, Kobe (JP); and Shinichiro Oguni, Kobe (JP)
Assigned to SYSMEX CORPORATION, Kobe (JP)
Filed by SYSMEX CORPORATION, Kobe (JP)
Filed on Jun. 29, 2020, as Appl. No. 16/915,851.
Application 16/915,851 is a division of application No. 14/061,333, filed on Oct. 23, 2013, abandoned.
Application 14/061,333 is a continuation of application No. PCT/JP2012/060432, filed on Apr. 18, 2012.
Claims priority of application No. 2011-101595 (JP), filed on Apr. 28, 2011.
Prior Publication US 2020/0326332 A1, Oct. 15, 2020
Int. Cl. G01N 33/50 (2006.01); G01N 15/14 (2006.01); G01N 15/1429 (2024.01); G01N 21/49 (2006.01); G01N 21/64 (2006.01); G01N 15/01 (2024.01)
CPC G01N 33/5094 (2013.01) [G01N 15/1429 (2013.01); G01N 15/147 (2013.01); G01N 21/49 (2013.01); G01N 21/6428 (2013.01); G01N 2015/011 (2024.01); G01N 2015/016 (2024.01)] 7 Claims
 
1. A method for determining whether a biological sample contains atypical lymphocytes or myeloblasts, the method comprising the steps of:
preparing a measurement sample by mixing the biological sample with a first reagent containing a fluorescent dye capable of staining nucleic acid and a second reagent containing cationic and nonionic surfactants and an aromatic organic acid at a concentration of not less than 30 mM and not more than 50 mM, the second reagent having pH of not lower than 5.5 and not higher than 7.0,
applying light with a flow cytometer to the prepared measurement sample and obtaining side-scattered light intensity and fluorescence intensity generated thereby,
preparing a scattergram comprising the obtained side-scattered light intensity and fluorescence intensity, the scattergram having two axes of side-scattered light intensity and fluorescence intensity,
classifying the monocytes and lymphocytes on the scattergram,
and identifying a cluster of atypical lymphocytes or a cluster of myeloblasts in the prepared measurement sample,
wherein a cluster of the atypical lymphocytes is shown to deviate towards a cluster of lymphocytes on the scattergram, and a cluster of the myeloblasts is shown to deviate towards a cluster of monocytes on the scattergram.