PMID- 28253379
OWN - NLM
STAT- MEDLINE
DCOM- 20180720
LR  - 20191210
IS  - 1560-2281 (Electronic)
IS  - 1083-3668 (Linking)
VI  - 22
IP  - 3
DP  - 2017 Mar 1
TI  - Considerations for point-of-care diagnostics: evaluation of acridine orange 
      staining and postprocessing methods for a three-part leukocyte differential test.
PG  - 35001
LID - 10.1117/1.JBO.22.3.035001 [doi]
AB  - There exists a broad range of techniques that can be used to classify and count 
      white blood cells in a point-of-care (POC) three-part leukocyte differential 
      test. Improvements in lenses, light sources, and cameras for image-based POC 
      systems have renewed interest in acridine orange (AO) as a contrast agent, 
      whereby subpopulations of leukocytes can be differentiated by colorimetric 
      analysis of AO fluorescence emission. We evaluated the effect on test accuracy 
      using different AO staining and postprocessing methods in the context of an 
      image-based POC colorimetric cell classification scheme. Thirty blood specimens 
      were measured for percent cell counts using our POC system and a conventional 
      hematology analyzer for comparison. Controlling the AO concentration used during 
      whole-blood staining, the incubation time with AO, and the colorimetric ratios 
      among the three population of leukocytes yielded a percent deviation of 0.706%, ? 
      1.534 % , and ? 0.645 % for the lymphocytes, monocytes, and granulocytes, 
      respectively. Overall, we demonstrated that a redshift in AO fluorescence was 
      observed at elevated AO concentrations, which lead to reproducible inaccuracy of 
      cell counts. This study demonstrates there is a need for a strict control of the 
      AO staining and postprocessing methods to improve test accuracy in these POC 
      systems.
FAU - Powless, Amy J
AU  - Powless AJ
AD  - University of Arkansas, Department of Biomedical Engineering, Fayetteville, 
      Arkansas, United States.
FAU - Conley, Roxanna J
AU  - Conley RJ
AD  - University of Arkansas, Pat Walker Health Center, Fayetteville, Arkansas, United 
      States.
FAU - Freeman, Karan A
AU  - Freeman KA
AD  - University of Arkansas, Pat Walker Health Center, Fayetteville, Arkansas, United 
      States.
FAU - Muldoon, Timothy J
AU  - Muldoon TJ
AD  - University of Arkansas, Department of Biomedical Engineering, Fayetteville, 
      Arkansas, United States.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PL  - United States
TA  - J Biomed Opt
JT  - Journal of biomedical optics
JID - 9605853
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*metabolism
MH  - Hematologic Tests/*standards
MH  - Leukocytes/*cytology
MH  - Point-of-Care Systems/standards
MH  - Reproducibility of Results
MH  - Staining and Labeling/standards
EDAT- 2017/03/03 06:00
MHDA- 2018/07/22 06:00
CRDT- 2017/03/03 06:00
PHST- 2016/12/01 00:00 [received]
PHST- 2017/02/08 00:00 [accepted]
PHST- 2017/03/03 06:00 [entrez]
PHST- 2017/03/03 06:00 [pubmed]
PHST- 2018/07/22 06:00 [medline]
AID - 2608663 [pii]
AID - 10.1117/1.JBO.22.3.035001 [doi]
PST - ppublish
SO  - J Biomed Opt. 2017 Mar 1;22(3):35001. doi: 10.1117/1.JBO.22.3.035001.