Reliable, Accurate Determination of the Leukocyte Differential of Leukopenic Samples by Using Hematoflow Method

Abstract

BackgroundHematology analyzers may ineffectively recognize abnormal cells, and manual differential counts may be imprecise for leukopenic samples. We evaluated the efficacy of the Hematoflow method for determining the leukocyte differential in leukopenic samples and compared this method with the manual differential method.MethodsWe selected 249 blood samples from 167 patients with leukopenia (WBC counts, 500-2,000/µL) for analysis in this study. The EDTA-anticoagulated blood samples were analyzed using an automatic blood cell counter (DxH800; Beckman Coulter, USA) and flow cytometry (FC 500; Beckman Coulter) by using Cytodiff reagent and analysis software (Beckman Coulter). Hematoflow results were selected or calculated from DxH800 and Cytodiff results. Two trained pathologists performed a manual differential count by counting 50-100 cells.ResultsThe precision of the Hematoflow method was superior to that of the manual method in counting 5 leukocyte subpopulations, immature granulocytes (IGs), and blasts. Blasts were detected in all 45 cases (100%) by Hematoflow. The correlation of the Cytodiff blast count to the reference count was high (r = 0.8325). For all other cell populations, the correlation of the Hematoflow results with the reference count was stronger than that of the other manual counts with the reference count.ConclusionsThe Hematoflow differential counting method is more reproducible and sensitive than manual counting, and is relatively easy to perform. In particular, this method detected leukemic blasts more sensitively than manual differential counts. The Hematoflow method is a very useful supplement to automated cell counting.

Evaluation of the leukocyte differential on a new automated flow cytometry hematology analyzer

“…We found a moderate correlation ( r = 0.5028) between the FC500 basophil count and reference counts. Some authors reported a low correlation between the Cytodiff basophil count and reference counts [8, 9], whereas some found a high correlation between these counts ( r = 0.8727) [6]. This is owing to CD2 + CRTH2 expressed in activated T cells, eosinophils, and basophils.…”

“…This is owing to CD2 + CRTH2 expressed in activated T cells, eosinophils, and basophils. Separation between basophils and non‐B‐non‐T blast populations was not effective in the Hematoflow method [6].…”

“…The leukocytes were differentiated into 16 cell populations (B-lymphocytes, CD16 ) T-lymphocytes, CD16 + T and NK cells, T and NK lymphocytes, total lymphocytes, CD16 monocytes, CD16 + monocytes, total monocytes, immature granulocytes [IGs], total eosinophils, mature neutrophils, total neutrophils, blasts B, blasts T, blasts non-B-non-T, and total basophils). Analysis procedures were conducted in accordance with the manufacturer's manual [6].…”

An extended leukocyte differential count (16 types of circulating leukocytes) using the cytodiff flow cytometric system can provide information for the discrimination of sepsis severity and prediction of outcome in sepsis patients

“…This system is based on the Beckman Coulter FC500 flow cytometer and enables the detection of a total of 16 types of circulating leukocytes [B lymphocytes, noncytotoxic T lymphocytes, cytotoxic T1NK (natural killer) lymphocytes, T1NK lymphocytes, total lymphocytes, CD (clusters of differentiation)16(-) monocytes, CD16(1) monocytes, total monocytes, immature granulocytes, eosinophils, mature neutrophils, total neutrophils, B cell blasts, T cell blasts, non-B and non-T cell blasts, and basophils] using a combination of six markers and five colors: CD36-FITC, CD2-PE, CRTH2-PE, CD19-ECD, CD16-PC5, and CD45-PC7 (2,3). This system is based on the Beckman Coulter FC500 flow cytometer and enables the detection of a total of 16 types of circulating leukocytes [B lymphocytes, noncytotoxic T lymphocytes, cytotoxic T1NK (natural killer) lymphocytes, T1NK lymphocytes, total lymphocytes, CD (clusters of differentiation)16(-) monocytes, CD16(1) monocytes, total monocytes, immature granulocytes, eosinophils, mature neutrophils, total neutrophils, B cell blasts, T cell blasts, non-B and non-T cell blasts, and basophils] using a combination of six markers and five colors: CD36-FITC, CD2-PE, CRTH2-PE, CD19-ECD, CD16-PC5, and CD45-PC7 (2,3).…”

A 5‐color flow cytometric method for extended 8‐part leukocyte differential

“…captodayonline.com/2015/ProductGuides). Only one of these methods offers a fully (10) automated WBC differential by automatic adjustment of the gates (11). This also has limitations as, for instance, lymphocyte subsets or blast cells lineage cannot be derived from morphology.…”