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Quality-Controlled Sputum Analysis by Flow Cytometry
Published on: August 9, 2021
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Computational Hyperspectral Microflow Cytometry
Hyo Geun Yun1, Yoel Alonso Cadierno2, Tae Won Kim1
1Department of Electronic Engineering, Hanyang University, Seoul, 04763, Republic of Korea.
Small (Weinheim an Der Bergstrasse, Germany)
|May 21, 2024
Summary
A new computational hyperspectral microflow cytometer (CHC) accurately distinguishes spectrally overlapping fluorophores in single cells. This breakthrough enhances portable cell analysis for diagnostics and immune cell monitoring.
Area of Science:
- Biomedical Engineering
- Optical Engineering
- Cell Biology
Background:
- Miniaturized flow cytometry is crucial for portable diagnostics and cell manufacturing monitoring.
- Current methods struggle to resolve spectrally overlapping fluorescence labels, limiting analytical precision.
- Accurate cell analysis requires advanced techniques for distinguishing specific cellular markers.
Purpose of the Study:
- To develop a computational hyperspectral microflow cytometer (CHC) for precise discrimination of spectrally overlapping fluorophores.
- To enhance the accuracy and reliability of microflow cytometry through advanced spectral detection and microfluidic focusing.
- To demonstrate the utility of CHC in analyzing T lymphocyte subpopulations and monitoring cell expansion.
Main Methods:
- Utilized a dispersive optical element and optimization algorithm to capture full fluorescence emission spectra from single cells.
- Achieved high spectral resolution (≈3 nm) in the visible range (450–650 nm).
- Integrated a microfluidic device with viscoelastic sheathless focusing for in-focus cell imaging.
Main Results:
- Successfully discriminated spectrally overlapping fluorophores labeling single cells with high accuracy.
- Demonstrated the capability of CHC to analyze T lymphocyte subpopulations.
- Showcased the monitoring of cellular composition changes during T cell expansion.
Conclusions:
- CHC represents a significant advancement in microflow cytometry, overcoming limitations in spectral resolution.
- The technology enables accurate analysis of complex cellular samples, including immune cells.
- CHC facilitates the broader application of microflow cytometry in diagnostics and bioprocess monitoring.
