US 9,810,618 B2
Basophil analysis system and method
Jiong Wu, Los Gatos, CA (US); Michael R. Buhl, San Ramon, CA (US); and Giacomo Vacca, San Jose, CA (US)
Assigned to Abbott Laboratories, Abbott Park, CA (US)
Filed by Abbott Laboratories, Abbott Park, IL (US)
Filed on Jul. 24, 2015, as Appl. No. 14/808,211.
Application 14/808,211 is a continuation of application No. 13/456,744, filed on Apr. 26, 2012, granted, now 9,091,625.
Claims priority of provisional application 61/482,549, filed on May 4, 2011.
Prior Publication US 2016/0018311 A1, Jan. 21, 2016
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 15/14 (2006.01); G01N 21/64 (2006.01); G01N 33/49 (2006.01); G01N 15/00 (2006.01); G01N 15/10 (2006.01)
CPC G01N 15/1434 (2013.01) [G01N 15/1459 (2013.01); G01N 21/6428 (2013.01); G01N 33/49 (2013.01); G01N 2015/008 (2013.01); G01N 2015/0069 (2013.01); G01N 2015/1006 (2013.01); G01N 2015/1402 (2013.01); G01N 2015/1477 (2013.01); G01N 2015/1486 (2013.01); G01N 2015/1488 (2013.01); G01N 2021/6439 (2013.01)] 18 Claims
1. A hematology analyzer for conducting a basophil analysis on a blood sample that contains a plurality of basophils and lymphocytes, the analyzer comprising:
an excitation source positioned to excite particles within the blood sample;
a plurality of detectors including (1) an axial light loss detector positioned to measure axial light loss from the excited blood sample, (2) an intermediate angle scatter detector positioned to measure intermediate angle scatter from the excited blood sample, (3) a side scatter detector positioned to measure 90° side scatter from the excited blood sample, and (4) a fluorescence detector positioned to measure fluorescence emitted from the excited blood sample;
a processor; and
a non-transitory computer-readable memory medium comprising instructions that when executed cause the processor to:
(a) dilute the blood sample with a reagent that includes a red blood cell (RBC) lysing agent and a cell membrane permeable, nucleic acid binding fluorescent dye;
(b) incubate the diluted blood sample of step (a) for an incubation period of time;
(c) deliver the incubated sample from step (b) to a flow cell in the hematology analyzer;
(d) excite the incubated sample from step (c) with an excitation source as the incubated sample traverses the flow cell;
(e) collect a plurality of light scatter signals and a fluorescence emission signal from the excited sample;
(f) prior to performing a basophil analysis, exclude nuclei-free events and retain nuclei-containing events using only a fluorescence trigger that is limited to fluorescence emission signals and is set to a fluorescence magnitude that is greater than fluorescence emission signals from RBCs, including RBC fragments, and is less than fluorescence emission signals from white blood cells (WBCs); and
(g) perform a basophil cluster analysis on the nuclei-containing events collected in step (f).