Noninvasive white blood cell quantification in umbilical cord blood collection bags with quantitative oblique back-illumination microscopy
Noninvasive white blood cell quantification in umbilical cord blood collection bags with quantitative oblique back-illumination microscopy
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DOI：
10.1111/trf.15704
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发表时间：
2020-02-14
期刊：
影响因子：
2.9
通讯作者：
Robles, Francisco E.
中科院分区：
文献类型：
--
作者：
Costa, Paloma Casteleiro;Ledwig, Patrick;Robles, Francisco E.
脐带血已成为治疗用造血干细胞和祖细胞的重要来源。然而，脐带血银行（CBB）努力解决与经济可行性相关的问题，部分原因是脐带血单位（CBUs）缺乏足够的总有核细胞用于储存或治疗用途，丢弃率高。目前，还没有可用的方法来评估CBB在收集点达到无创储存标准的可能性，这将提高CBB的效率和经济可行性。为了克服这一限制，我们应用了一种新的无标签光学成像方法，称为定量斜背照明显微镜（qOBM），该方法可以产生层析相位和吸收对比，以成像采集袋内的血液。开发了一种自动分割算法来计数白细胞和红细胞（rbc）并评估红细胞压积。测量了15个CBUs。结果qOBM明显区分红细胞和有核细胞。与血液学分析相比，细胞计数分析的平均误差为13%，具有近乎完美的一对一关系（斜率= 0.94）和强相关系数（r = 0.86）。评估红细胞压积的初步结果也显示与期望值非常吻合。每个CBU成像具有统计学意义的细胞数量的采集时间约为1分钟。结论qOBM法对采集袋内血液的定量具有较好的效果。由于该方法是自动化和快速的，它可以在几分钟内对CBUs进行量化，而不会破坏CBUs的无菌环境。qOBM可以通过避免最终不符合存储标准的CBB的处理费用来降低CBB的成本。
BACKGROUND Umbilical cord blood has become an important source of hematopoietic stem and progenitor cells for therapeutic applications. However, cord blood banking (CBB) grapples with issues related to economic viability, partially due to high discard rates of cord blood units (CBUs) that lack sufficient total nucleated cells for storage or therapeutic use. Currently, there are no methods available to assess the likelihood of CBUs meeting storage criteria noninvasively at the collection site, which would improve CBB efficiency and economic viability.MATERIALS AND METHODS To overcome this limitation, we apply a novel label-free optical imaging method, called quantitative oblique back-illumination microscopy (qOBM), which yields tomographic phase and absorption contrast to image blood inside collection bags. An automated segmentation algorithm was developed to count white blood cells and red blood cells (RBCs) and assess hematocrit. Fifteen CBUs were measured.RESULTS qOBM clearly differentiates between RBCs and nucleated cells. The cell-counting analysis shows an average error of 13% compared to hematology analysis, with a near-perfect, one-to-one relationship (slope = 0.94) and strong correlation coefficient (r = 0.86). Preliminary results to assess hematocrit also show excellent agreement with expected values. Acquisition times to image a statistically significant number of cells per CBU were approximately 1 minute.CONCLUSION qOBM exhibits robust performance for quantifying blood inside collection bags. Because the approach is automated and fast, it can potentially quantify CBUs within minutes of collection, without breaching the CBUs' sterile environment. qOBM can reduce costs in CBB by avoiding processing expenses of CBUs that ultimately do not meet storage criteria.