Key Laboratory of Precision Scientific Instrumentation of Anhui Higher Education Institutes, Department of Precision Machinery and Precision Instrumentation, University of Science and Technology of China, 96 Jinzhai Road, Hefei, 230027, Anhui, China.
Hefei National Laboratory for Physical Sciences at the Microscale, University of Science and Technology of China, 96 Jinzhai Road, Hefei, 230027, Anhui, China.
Anal Bioanal Chem. 2021 Aug;413(20):5025-5035. doi: 10.1007/s00216-021-03466-6. Epub 2021 Jun 25.
While many clinical laboratory tests are now highly automated, body fluid cell counting, particularly in low-cellularity samples such as cerebral spinal fluid (CSF), is often performed manually. Here, we report a simple, cost-effective method to obtain white and red blood cell counts from human body fluids such as CSF. The method consists of a compact, automated, and low-cost fluorescence microscope system, coupled to a sample chamber containing all of the necessary reagents in dry form to stain and prepare the sample. Sample focus and scanning are handled automatically, and the acquired multimodal images are automatically analyzed to extract cell counts. Comparison with manual counting on over 200 clinical samples shows excellent agreement. As the system counts a substantially larger image region than a standard manual cell count, we find our sensitivity to extremely low cellularity samples to potentially be higher than the manual gold standard, evidenced by our system recording images of cells in samples whose cell count was registered as "0" by a trained user. Thus, our system holds promise for routine, automated, and sensitive analysis of body fluids whose cellularity extends across a wide dynamic range.
虽然现在许多临床实验室测试已经高度自动化，但体液细胞计数，特别是在脑脊液（CSF）等低细胞密度样本中，通常仍需要手动进行。在这里，我们报告了一种简单、经济有效的方法，可从人体体液（如 CSF）中获得白细胞和红细胞计数。该方法包括一个紧凑、自动化且成本低廉的荧光显微镜系统，与一个包含所有必要试剂的干燥形式的样本腔耦合，用于对样本进行染色和准备。样本聚焦和扫描自动处理，获取的多模态图像自动分析以提取细胞计数。与超过 200 个临床样本的手动计数进行比较，结果显示出极好的一致性。由于该系统计数的图像区域比标准的手动细胞计数大得多，因此我们发现，与手动金标准相比，我们对极低细胞密度样本的灵敏度可能更高，这一点可以从我们的系统记录下训练有素的用户登记为“0”的细胞计数的样本中的细胞图像中得到证明。因此，我们的系统有望实现常规、自动化和对细胞密度广泛动态范围的体液的敏感分析。
