Differential Leukocyte Counting via Fluorescent Detection and Image Processing on a Centrifugal Microfluidic Platform
| Autor: | Max L. Balter, Alvin I. Chen, Tim Maguire, Brian Bixon, Alexander Gorshkov, Alexander Fromholtz, Martin L. Yarmush, Vincent Martin, C. Amara Colinco |
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| Rok vydání: | 2017 |
| Předmět: |
Pixel
General Chemical Engineering Monocyte 010401 analytical chemistry Microfluidics General Engineering Analytical chemistry Image processing 02 engineering and technology Biology 021001 nanoscience & nanotechnology 01 natural sciences Sample (graphics) Fluorescence Article 0104 chemical sciences Analytical Chemistry medicine.anatomical_structure White Cell medicine Fluorescence microscope 0210 nano-technology Biomedical engineering |
| Zdroj: | Analytical methods : advancing methods and applications. 8(47) |
| ISSN: | 1759-9660 |
| Popis: | Centrifugal microfluidics has received much attention in the last decade for the automation of blood testing at the point-of-care, specifically for the detection of chemistries, proteins, and nucleic acids. However, the detection of common blood cells on-disc, particularly leukocytes, remains a challenge. In this paper, we present two analytical methods for enumerating leukocytes on a centrifugal platform using a custom-built fluorescent microscope, acridine orange nuclear staining, and image processing techniques. In the first method, cell analysis is performed in glass capillary tubes; in the second, acrylic chips are used. A bulk-cell analysis approach is implemented in both cases where the pixel areas of fractionated lymphocyte/monocyte and granulocyte layers are correlated with cell counts. Generating standard curves using porcine blood sample controls, we observed strong linear fits to measured cell counts using both methods. Analyzing the pixel intensities of the fluorescing white cell region, we are able to differentiate lymphocytes from monocytes via pixel clustering, demonstrating the capacity to perform a 3-part differential. Finally, a discussion of pros and cons of the bulk-cell analysis approach concludes the paper. |
| Databáze: | OpenAIRE |
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