A fixed cytometer chip for identification of cell populations and real-time monitoring of single-cell apoptosis under gradient UV radiation
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Cytometry is a basic method to determine cell populations and morphology. Flow cytometry and hemocytometry are the two most common methods among cytometric technologies. However, flow cytometry needs bulky and expensive equipment as well as professional operations, while hemocytometry is limited by its simple function. Both of them are not suitable for real-time monitoring of the morphological changes of single cells. Here, we developed a fixed cytometer chip with two functional modes for both identification of cell populations (I-mode) and real-time monitoring of single-cell morphological changes (M-mode). In I-mode, the fixed cytometer chip was employed to evaluate the cell populations, the results were in accordance with those from the hemocytometer counting and flow cytometry. Besides that, the cell populations were further precisely identified by measuring two-color fluorescence intensities of single cells, which were consistent with the dual parameter analysis in flow cytometry. In M-mode, the chip was applied to real-time monitoring of the single-cell apoptosis under gradient UV radiation, generated by a novel stair-like UV shield. The dynamic apoptotic morphologies of a large number of single cells were monitored in real-time by time-lapse imaging. In addition, we integrated eight parallel channels on a 60 mm × 30 mm chip, and the chip could achieve scalable single-cell capture and analysis capability. This fixed cytometer chip is bifunctional, easy-to-handle, universal, and scalable.
KeywordsCytometer chip Cell population identification Real-time monitoring Cell Apoptosis Gradient UV radiation
We gratefully acknowledge the financial support from the National Natural Science Foundation of China (81771968, 81472842, 21778071, and 31400087), Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support (20181705), Shanghai Talent Development Fund (2017053), Translational Medicine Cross Research Grant of Shanghai Jiao Tong University (ZH2018ZDA05), Youth Innovation Promotion Association CAS and Suzhou Institute of Nano-Tech and Nano-Bionics Owned Fund (Y5AAS11001).
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
- Del Carratore R, Della Croce C, Simili M, Taccini E, Scavuzzo M, Sbrana S (2002) Cell cycle and morphological alterations as indicative of apoptosis promoted by UV irradiation in S-cerevisiae. Mut Res Gen Toxicol Environ 513:183–191. https://doi.org/10.1016/s1383-5718(01)00310-2 CrossRefGoogle Scholar
- Pierzchalski A, Mittag A, Tarnok A (2011) Introduction a: recent advances in cytometry instrumentation, probes, and methods-review. In: Darzynkiewicz Z, Holden E, Orfao A, Telford W, Wlodkowic D (eds) Recent advances in cytometry, part a: instrumentation, methods, Fifth Edition, vol 102. Methods in Cell Biology. pp 1–21. https://doi.org/10.1016/b978-0-12-374912-3.00001-8 Google Scholar
- Rawstron AC, de Tute RM, Haughton J, Owen RG (2016) Measuring disease levels in myeloma using flow cytometry in combination with other laboratory techniques: lessons from the past 20 years at the leeds haematological malignancy diagnostic service. Cytom Part B Clin Cytom 90:54–60. https://doi.org/10.1002/cyto.b.21271 CrossRefGoogle Scholar