Correlation between the cell population in the automated hematology analyzer high‐fluorescence region and atypical lymphocyte flags

Introduction

During routine blood measurements using an automated hematology analyzer, two easily confused types of suspect flags related to lymphocytes often appear: atypical and immature lymphocytes. The aim of this study was to investigate the correlation of high fluorescence cell (HFC) parameter and lymphocyte flags determined from an automated hematology analyzer.

Methods

A total of 93 patients affected by various pathologic conditions (viral infection, immunological disease, oncological disease and tumor) were divided into an “atypical lymphocytes” group (“atypical” for short), an “immature lymphocytes/blasts” flag group (abnormal), a mixed‐flag group that includes “atypical lymphocytes” (mixed), and a non‐flag group (non‐flag).

Results

The numbers of HFCs in the atypical, abnormal, mixed, and non‐flag groups were 1.8% (0.9%‐5.5%), 0.7% (0.1%‐5.0%), 2.3% (1.2%‐5.0%), and 0.8% (0.7%‐1.2%), respectively. The HFCs of “atypical” appeared as a separate cluster with clear boundaries. The HFCs of “abnormal” as an unclear boundaries, and it was difficult to accurately distinguish between the HFCs from the immature lymphocytes and the normal lymphocytes. The lower limit of HFC when the atypical lymphocyte flag appeared was 0.04 × 10⁹/L. The number of HFCs was similar to atypical lymphocytes detected by microscopy and CD19⁺CD20⁻CD27⁺⁺ cells by flow cytometry at 78% and 76%, respectively. The number of HFCs detected in “atypical” and CD19⁺CD20⁻CD27⁺⁺ cells showed good consistency (r = .715), whereas the consistency was poorest for “abnormal” (r = .176).

Conclusion

It demonstrates that HFCs reflects atypical lymphocytes better than immature lymphocytes/blasts.

     

Application of Pop-off Method to Bone Marrow and Peripheral Blood Specimens for Purposes of Electron Microscopy

In this paper, we describe a pop-off method applicable to the hematological field. Bone marrow or peripheral blood specimens from patients were placed on a clean glass slide and fixed immediately in 2% glutaraldehyde solution for 10 minutes. For the DAB reaction, the slide was immersed in the DAB reagent for 30 minutes, and post fixed with 1 % OsO₄ solution for 1 hour. Specimens on the slide were washed with buffer solution, dehydrated and polymerized directly on the slide. A gelatin capsule filled with Epon mixed monomer was then reversed over the specimen. After polymerization was completed, the specimen was popped off from the slide to the capsule and trimmed carefully to prepare for ultrathin sectioning. This method allows the entire sequence of tissue preparation to be carried out on the slide, from fixation to embedding, and even, especially in scanty specimens, including the DAB reaction. Electron microscopic findings in specimens prepared by this technique show excellent preservation and the absence of specific artifacts.     

Primary dural lymphoma: A novel concept of heterogeneous disease

Spinal primary dural lymphoma (PDL) is uncommon with a total of 37 previous well‐documented cases reported, including one diagnosed in the authors' institution. More recently we encountered an additional case of spinal PDL that, similarly to our previous case, was grade 1–2 follicular B‐cell PDL. Our two cases were diagnosed over a 3‐year interval in a 72‐year‐old female and a 74‐year‐old male, respectively. An exhaustive literature review on PDL was performed consequently to reveal that: (i) spinal and cerebral sites of involvement by PDL are constantly mutually exclusive; and (ii) unlike cerebral PDL, which is usually of marginal zone B‐cell type, only two of the 38 cases of spinal PDL were diagnosed as such, diffuse large B‐cell lymphoma being the most commonly encountered type in the spine. This divergence infers that, in contrast to the prevailing concept that PDL is a unique disease group, PDL appears to be rather heterogeneous with a difference in predilection of lymphoma type for the anatomical site of dural involvement. Such a site‐specific lymphoma‐type predilection phenomenon, well‐recognized in other organ systems, has not been acknowledged in PDL. This report brings new insights into PDL, and may contribute to a better understanding of nervous system pathophysiology and lymphoma classification.     

XN-1000希森美康对孕妇外周静脉血分析提示与细胞涂片染色分析的相关性

目的探讨XN-1000希森美康细胞分析仪对孕妇外周静脉血白细胞分类计数、有核红细胞计数分析提示与相关细胞涂片染色人工分析结果相关性及意义。方法采用XN-1000型希森美康细胞分析仪对妇产科采集的280份外周静脉血进行白细胞分类计数、有核红细胞计数分析,同时对应外周静脉血标本进行细胞涂片染色分析,比较仪器分析和细胞染色人工分析结果。结果仪器分析重复性良好,仪器分析和细胞染色人工分析具有较高的相关性,报警提示结果和细胞染色人工分析阳性结果特异性大于95%,灵敏度100%,阳性预测值大于90%,阴性预测值100.00%。结论 XN-1000希森美康细胞分析仪分析报警提示可以有效筛选阳性标本,同时对阳性标本需要细胞染色人工分析,避免假阳性结果。