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A smart preparation strategy for point-of-care cellular counting of trace volumes of human blood

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Abstract

Blood counting is one of the most commonly ordered clinical assays, and is often part of the basis for initial diagnosis and screening for disease. While substantial prior research has shown the ability of portable instruments to accurately produce blood counts through image- or flow-based cytometry, these methods require complex sample preparation using either costly commercial imaging chambers or complicated reagents. To address these issues, in this paper we developed a method to prepare trace volumes of whole blood aimed at portable blood counting. The strategy is based on pre-storing dry-form reagents and fabricating a specifically designed cell counter. In order to obtain total cell counts for red blood cells, platelets, and 3-part differentials of white blood cells, two parallel counting chambers with different depths are made from cost- and environmentally friendly materials using soft lithography. As little as 1 μl of whole blood is prepared with pre-stored reagents in centrifuge vials, whereas red blood cells are sphered and white blood cells are stained at the same time. Driven by the capillary force, prepared blood samples enter the hydrophilic chambers automatically. Monolayers of cells are formed when the blood dilution factors and the chamber depths are co-optimized. Combined with our previous custom-built instrument and automated analysis algorithm, the sample preparation strategy allows producing counting results with excellent agreement to a gold-standard clinical hematology instrument. The success of this preparation method may further advance applications of our technology for global use in low-resource settings where central hematology laboratories are not accessible.

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Funding

This work is supported by the National Natural Science Foundation of China (21605054), Central China Normal University (CCNU) new PI start-up fund (210-31102), and self-determined research funds of CCNU (234-20205016002, 234-20205170355) from the colleges’ basic research and operation of Ministry of Education (MOE). TG acknowledges support from the Program of Introducing Talents of Discipline to University of China (111 program, B17019), and the CCNU Program of Innovation and Entrepreneurship for college students (220-20205180419). ZJS acknowledges support from the 1000 Young Talents Global Recruitment Plan, and from the Ministry of Science and Technology of the People’s Republic of China (2016YFA020130).

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Author notes

  1. Xinyi Li and Qiong Deng contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Pesticide and Chemical Biology of Ministry of Education, and Chemical Biology Center, College of Chemistry, Central China Normal University, Wuhan, 430079, Hubei, China

    Xinyi Li, Qiong Deng, Yangfei Chen, Yuchen Tang & Tingjuan Gao

  2. State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, 430071, Hubei, China

    Hongping Liu

  3. Prenatal Diagnostics Center, Jieyang Women and Children’s Health Care Hospital, Jieyang, 522000, Guangdong, China

    Youwang Lei

  4. Clinical Laboratory Center, Wuhan No. 7 Hospital, Wuhan, 430071, Hubei, China

    Pengwei Fan & Bin Wang

  5. Department of Precision Machinery and Precision Instrumentation, University of Science and Technology of China, Hefei, 230027, Anhui, China

    Zachary J. Smith

Authors
  1. Xinyi Li
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  2. Qiong Deng
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  3. Hongping Liu
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  4. Youwang Lei
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  8. Zachary J. Smith
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  9. Yuchen Tang
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  10. Tingjuan Gao
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Correspondence to Yuchen Tang or Tingjuan Gao.

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Li, X., Deng, Q., Liu, H. et al. A smart preparation strategy for point-of-care cellular counting of trace volumes of human blood. Anal Bioanal Chem 411, 2767–2780 (2019). https://doi.org/10.1007/s00216-019-01738-w

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  • DOI: https://doi.org/10.1007/s00216-019-01738-w

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