Preserving whole blood in formalin extends the specimen stability period for manual cell counts for fish
Preserving whole blood in formalin extends the specimen stability period for manual cell counts for fish
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DOI：
10.1111/vcp.12214
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发表时间：
2014-12-01
影响因子：
1.2
通讯作者：
Rosemary, Kevin
中科院分区：
文献类型：
--
作者：
Arnold, Jill E.;Matsche, Mark A.;Rosemary, Kevin
背景：鱼类的总血细胞计数具有挑战性，特别是由于细胞脆弱。收集后数小时内可能会发生血小板聚集、细胞变形和裂解；因此，血细胞计数器可能无法对大量样本进行计数。在福尔马林中保存全血提供了一种延长细胞计数样本稳定期的简单方法。 目的：本研究的目的是使用新鲜采集和福尔马林固定的样本来表征和比较 Natt-Herrick 方法对鱼类血细胞计数的分析性能。方法：比较来自 11 个软骨鱼物种 (n = 36) 的样本的 WBC 计数。对 50 条条纹鲈 (Morone saxatilis) 的白细胞、红细胞和血小板计数进行了比较，包括福尔马林保存细胞的 2 个时间点（大约 1 周和 1 个月储存）。计算变异系数 (CV)、偏差和总误差 (TEcalc)。将 TEcalc 与临床实验室改进修正案 (CLIA) 定义的人类血细胞计数的允许总误差 (TEa) 进行比较。结果：新鲜和固定的 WBC 和血小板计数的 CV 和 TEcalc 符合 CLIA 限值。在新鲜和保存的细胞中，RBC CV 和 TEcalc 均比 TEa 高出近 3 倍。结论：将新鲜采集的血液保存在 10% 福尔马林中是一种可靠的方法，可在采集后长达 1 个月内保持手动计数的细胞形态。这对于实验室访问受到限制的现场研究特别有用。需要进一步评估以确定手动红细胞计数在鱼类中的临床用途。
Background: Total blood cell counts for fish are challenging, especially due to cell fragility. Thrombocyte aggregation, cell distortion, and lysis can occur within hours of collection; therefore, hemocytometer counts may not be possible for large sample numbers. Preserving whole blood in formalin offers a simple way to extend the specimen stability period for cell counts.Objectives: The objectives of this study were to characterize and compare the analytical performance of the Natt-Herrick method for fish blood cell counts using freshly collected and formalin-fixed specimens.Methods: Specimens from 11 elasmobranch species (n = 36) were compared for WBC counts. WBC, RBC, and thrombocyte counts from 50 striped bass, Morone saxatilis, were compared including 2 time points for the formalin-preserved cells (appr. 1-week and 1-month storage). Coefficient of variation (CV), bias, and total error (TEcalc) were calculated. TEcalc was compared with allowable total error (TEa) defined by the Clinical Laboratory Improvements Amendment (CLIA) for human blood cell counts.Results: The CV and TEcalc for fresh and fixed WBC and thrombocyte counts met CLIA limits. In both the fresh and preserved cells, the RBC CV and TEcalc were nearly 3 times higher than the TEa.Conclusions: Preserving freshly collected blood in 10% formalin is a reliable method to maintain cell morphology for manual counts for up to 1 month post collection. This is especially useful for field studies, where laboratory access is limited. Further evaluation is needed to determine the clinical usefulness of the manual RBC count in fish.