Citation: MAO Cui-Ping, LIN Yao, NIU Qian, XUE Cheng-Feng, YAN Xiao-Mei. Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(7): 838-846. doi: 10.19756/j.issn.0253-3820.201227 shu

Development and Applications of A Highly Sensitive and Wide Dynamic Range Nano-Flow Cytometer

Department of Chemical Biology, The MOE Key Laboratory of Spectrochemical Analysis&Instrumentation, The Key Laboratory for Chemical Biology of Fujian Province, Collaborative Innovation Center of Chemistry for Energy Materials, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China

Received Date: 24 April 2020
Revised Date: 14 May 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21627811, 21934004).

Nanoparticles (NPs) are playing increasingly important roles in the fields of biomedicine, biochemical analysis, and energy materials. Because NPs are intrinsically heterogeneous, rapid and high resolution size distribution measurement of NPs is a challenging task. Compared with traditional approaches for the analysis of NPs, such as electron microscopy, atomic force microscopy, dynamic light scattering, and nanoparticle tracking analysis, flow cytometry is a well-established technique for high-throughput, quantitative, and multi-parameter analysis of individual cells and microscopic particles. However, it has been extremely difficult for conventional flow cytometry to detect polystyrene NPs smaller than 200 nm in diameter via light scattering. Employing strategies for single molecule fluorescence detection in a sheathed flow, we have recently developed Nano-flow cytometer (nFCM) that enables light scattering detection of single silica nanoparticles (SiO₂ NPs) as small as 24 nm in diameter. However, the dynamic range of the nFCM is limited by the maximum photon count of the single photon counting avalanche photodiode (APD) detector. Owing to the exponential dependence of scattered light intensity on particle size, there is an urgent need to enhance the dynamic range of the nFCM for the analysis of samples with wide size distributions. Adopting strategies of beam shaping with binary optical element, background reduction upon the combined usage of an iris-diaphragm and a rectangular aperture, and signal enhancement using high quantum efficiency PMT, here we reported the development of a highly sensitive and wide dynamic range nFCM. Sensitive light-scattering detection of single SiO₂ NPs as small as 59 nm in diameter was achieved with a signal-to-noise ratio of 119. Baseline separation of seven different sizes of SiO₂ NPs ranging from 59 to 222 nm was obtained. The as-developed nFCM was successfully applied to the size distribution analysis of outer membrane vesicles derived from Salmonella Typhimurium.
- Single-nanoparticle detection,
- Light scattering,
- Size distribution,
- Nano-flow cytometry,
- Bacterial outer membrane vesicles
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