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Comparative Study
. 2018 Apr 26;13(4):e0195923.
doi: 10.1371/journal.pone.0195923. eCollection 2018.

Evaluation of Cell Count and Classification Capabilities in Body Fluids Using a Fully Automated Sysmex XN Equipped With High-Sensitive Analysis (hsA) Mode and DI-60 Hematology Analyzer System

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Free PMC article
Comparative Study

Evaluation of Cell Count and Classification Capabilities in Body Fluids Using a Fully Automated Sysmex XN Equipped With High-Sensitive Analysis (hsA) Mode and DI-60 Hematology Analyzer System

Hiroyuki Takemura et al. PLoS One. .
Free PMC article

Abstract

The XN series automated hematology analyzer has been equipped with a body fluid (BF) mode to count and differentiate leukocytes in BF samples including cerebrospinal fluid (CSF). However, its diagnostic accuracy is not reliable for CSF samples with low cell concentration at the border between normal and pathologic level. To overcome this limitation, a new flow cytometry-based technology, termed "high sensitive analysis (hsA) mode," has been developed. In addition, the XN series analyzer has been equipped with the automated digital cell imaging analyzer DI-60 to classify cell morphology including normal leukocytes differential and abnormal malignant cells detection. Using various BF samples, we evaluated the performance of the XN-hsA mode and DI-60 compared to manual microscopic examination. The reproducibility of the XN-hsA mode showed good results in samples with low cell densities (coefficient of variation; % CV: 7.8% for 6 cells/μL). The linearity of the XN-hsA mode was established up to 938 cells/μL. The cell number obtained using the XN-hsA mode correlated highly with the corresponding microscopic examination. Good correlation was also observed between the DI-60 analyses and manual microscopic classification for all leukocyte types, except monocytes. In conclusion, the combined use of cell counting with the XN-hsA mode and automated morphological analyses using the DI-60 mode is potentially useful for the automated analysis of BF cells.

Conflict of interest statement

Competing Interests: The Department of Next Generation of Hematology Laboratory Medicine at Juntendo University Graduate School of Medicine has been supported by Sysmex. KK, KU and AK are employees of Sysmex. The study was performed as a collaborative study by scientifically proper methods without any bias. The participation of these authors does not alter our adherence to PLOS ONE policies on sharing data and materials.

Figures

Fig 1
Fig 1. Coefficient of variations (% CV) measured with the XN-hsA and XN-BF modes as a function of CSF sample cell density.
The right panel shows the magnified image of the portion of the plot (in the red rectangle).
Fig 2
Fig 2. Plots of cell counts against the theoretical cell densities using serially diluted body fluid samples measured with the XN-hsA and XN-BF modes.
Fig 3
Fig 3. Correlation of cell density analyses between the XN-hsA and XN-BF modes and manual microscopic examination using CSF samples (n = 60).
The raw data are shown in S1 Table.
Fig 4
Fig 4
(A) Comparison of the classifications of four different leukocyte types using DI-60 and manual microscopic examination in CSF samples (n = 34). Neut depicts neutrophils; Lymph, lymphocytes; Eo, eosinophils; Mo, monocytes. (B) Classifications of four different leukocyte types using the DI-60 and manual microscopic examination in other body fluid samples (n = 60). The raw data are shown in S2 Table.

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Grant support

This work was supported in part by a Grant-in-Aid for Scientific Research (C), Japan, a Grant-in-Aid (S1311011) from the Foundation of Strategic Research Projects in Private Universities from the MEXT, Japan (to YT). The Department of Next Generation of Hematology Laboratory Medicine at Juntendo University Graduate School of Medicine has been partially supported by Sysmex. KK, KU and AK are employees of Sysmex. The study was performed as a collaborative study by scientifically proper methods without any bias. The participation of these authors does not alter our adherence to PLOS ONE policies on sharing data and materials. We also state that the funder provided support in the form of salaries to authors [KK, AK, and KU], but did not have any judgmental roles in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are described in the ‘author contributions’ section.
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