Application of flow cytometry for the rapid determination of erythrocyte osmotic fragility

doi:10.3969/j.issn.1673-8640.2016.010.017

Abstract

Abstract:

Objective To establish a rapid determination of erythrocyte osmotic fragility by flow cytometry,and to evaluate the feasibility of determining erythrocyte osmotic fragility by flow cytometry. Methods A total of 30 anticoagulation peripheral blood specimens were collected,and each specimen was divided into 2 parts,one group collected within 2 h as normal control group and another group incubated at 37 ℃ for 24 h as positive control group. A total of 3 patients diagnosed as hereditary spherocytosis were enrolled as disease group. The erythrocyte osmotic fragility was determined by flow cytometry. The residue percentage of red blood cells was analyzed by flow cytometry. The optimal volume ratio of normal saline and sterile water was identified. The influence of ethylene diamine tetraacetic acid（EDTA） and heparin anticoagulants was evaluated. A total of 14 cases of heparin anticoagulation specimens were collected,and the red blood cell residue percentage was determined by improved Sanford and flow cytometry simultaneously. The characterization of determining erythrocyte osmotic fragility by flow cytometry and improved Sanford was evaluated.Results By flow cytometry,red blood cell residue percentages were 72.03% ± 5.68% in normal control group and 46.47% ± 15.83% in positive control group（P<0.05）. The disease group showed a significant reduction in red blood cell residue percentage（5.76% ± 3.54%） compared to that in normal control group（P<0.001）. When the volume ratio of normal saline to sterile water was 1.2∶0.8,there was significant difference of red blood cell residue percentage between the 2 groups（P<0.05）. Compared with EDTA anticoagulant,heparin anticoagulant was more suitable for the determination of erythrocyte osmotic fragility. There was a good consistency between flow cytometry and improved Sanford in the determination of erythrocyte osmotic fragility,but flow cytometry had better sensitivity than improved Sanford. Conclusions Flow cytometry has the characteristics of operation simplicity and sensitivity,and has a practical application feasibility in the determination of erythrocyte osmotic fragility.

Key words: Erythrocyte osmotic fragility, Flow cytometry, Sanford

CLC Number:

R446.1

WANG Weiwei, YUAN Xiangliang, FEI Qili, SHEN Lisong. Application of flow cytometry for the rapid determination of erythrocyte osmotic fragility[J]. Laboratory Medicine, 2016, 31(10): 911-916.

Figures/Tables 7

References 16

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项目	1	2	3	4	5	6	7	8	9	10	11	12	13
9 g/L NaCl（mL）	4.25	3.75	3.50	3.25	3.00	2.75	2.50	2.25	2.00	1.75	1.50	1.00	0.50
无菌水（mL）	0.75	1.25	1.50	1.75	2.00	2.25	2.50	2.75	3.00	3.25	3.50	4.00	4.50
NaCl（g/L）	8.50	7.50	7.00	6.50	6.00	5.50	5.00	4.50	4.00	3.50	3.00	2.00	1.00

项目	1	2	3	4	5	6	7	8	9	10	11	12	13
9 g/L NaCl（mL）	4.25	3.75	3.50	3.25	3.00	2.75	2.50	2.25	2.00	1.75	1.50	1.00	0.50
无菌水（mL）	0.75	1.25	1.50	1.75	2.00	2.25	2.50	2.75	3.00	3.25	3.50	4.00	4.50
NaCl（g/L）	8.50	7.50	7.00	6.50	6.00	5.50	5.00	4.50	4.00	3.50	3.00	2.00	1.00