Three levels of internal quality control (IQC) materials were regularly evaluated for all routine hematologic parameters. The characteristic that distinguishes HFLCs from other cells is the high intensity of their fluorescent signal, which reflects an abundance of RNA content in HFLCs. HFLCs are located separately from monocytes and lymphocytes on the scattergram that is generated by the analyzer. HFLCs were analyzed in the WDF channel of the analyzer using polymethine fluorescent dye, with analysis based on the flow cytometry principle. The analyzer was operated in routine mode according to the manufacturer-provided instructions. At least 3 mL of blood was mixed thoroughly before testing.
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Only 1 selected Sysmex XN module was assessed. This unit comprises 1 sampler, 2 Sysmex XN modular analyzers, and 1 Sysmex SP-10 slide maker and stainer (Sysmex Corporation). We used the Sysmex XN-3000 (Sysmex Corporation) to analyze blood count and HFLC in this study. HFLC Count by Automated Hematology Analyzer Diagnosis of infectious diseases required positive pathogen isolation, serologic testing, or specific antigen detection. After blood analysis was completed for each specimen, the principle diagnoses were reviewed and categorized into subgroups (ie, infectious diseases, immunological disorders, malignancies, and others). The investigators were masked to CBC parameters and patient data examinations included blood smears. After automated analysis, specimens were further examined for microscopic AL count. Complete blood count (CBC), HFLC analysis, and blood-smear preparation were performed within 4 hours after blood collection. Dipotassium ethylenediaminetetraacetic acid (EDTA) tubes (VACUETTE Greiner Bio-One International GmbH) were used for blood collection. These specimens were sent from inpatient and outpatient departments to the central laboratory at the Department of Clinical Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. We included a total of 320 blood specimens with various levels of HFLC from 0 to 3.4 × 10 9 per L in the study. Further, the clinical significance of HFLC was determined. Accordingly, the aim of this study was to investigate association between automated HFLC and AL counts.
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To our knowledge, no study has specifically investigated agreement between HFLC and manual microscopic AL counts, and the clinical significance of HFLC count.
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Because AL is a reactive form of lymphocyte that can be stimulated by immune processes, the number of HFLCs may also correlate with that of AL. Previous studies 5, 6 demonstrated the correlation between HFLCs and activated B lymphocytes, and between HFLCs and plasma cells in peripheral blood. HFLC count reveals a cell population that can be distinguished from monocytes and normal lymphocytes in a 5-part differential channel by the higher intensity of their fluorescent signals. High fluorescence lymphocyte cell (HFLC) count is a hematologic research marker that can be analyzed using an automated instrument. 4 Manual AL counting is also time- and labor-intensive. However, manual AL counting requires examiner expertise to produce an accurate result variability in results may occur among examiners due to examiner subjectivity. This manual counting method is widely used in most clinical laboratories because blood smear preparation is inexpensive and easy to perform.
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Then, a manual microscopic count is performed to estimate the AL count. Most routinely used automated hematology analyzers cannot perform an accurate AL count however, they are designed to generate a flag to alert laboratory staff to the presence of ALs in a specimen. 2 As a result of these variations, accurate enumeration of ALs can be challenging in routine clinical practice. Several AL subtypes have been described, including Downey type I, Downey type II, Downey type III, and plasmacytoid lymphocyte. Characteristics of one AL may differ from those of other ALs in the same blood smear. ALs are morphologically distinct from mature lymphocytes by their heterogeneity in size and shape. 1–3 Identification of peripheral blood ALs is normally used to support the diagnosis of viral infection. High-fluorescent lymphocyte cell, atypical lymphocyte, hematology analyzer, instrument evaluation, dengue infection, Sysmex XN-3000Ītypical lymphocyte (AL), or reactive lymphocyte, plays a role in immune response to various conditions, including infections, drug hypersensitivity, immunological disorders, and lymphoproliferative disorders.