研究目的
To evaluate the feasibility and limitations of lifetime encoding in flow cytometry with a compact setup and straightforward time-domain measurements utilizing lifetime-encoded beads.
研究成果
The study concludes that the presented LT-FCM platform allows for the discrimination of different lifetime codes, with the current performance level for lifetime encoding using a single excitation wavelength and a single detection channel being five codes involving organic and inorganic fluorophores. Discrimination capabilities could be further improved by technical modifications.
研究不足
The study acknowledges the need for technical modifications to improve discrimination capabilities, such as employing detectors with better dynamic range or flexible adaption of the measurement time window for each object, as well as software upgrades.
1：Experimental Design and Method Selection:
The study involved the development of a flow cytometry platform with time-resolved detection based on a compact setup and straightforward time-domain measurements.
2：Sample Selection and Data Sources:
Lifetime-encoded luminescent beads loaded with different organic fluorophores and semiconductor quantum dots were used as model systems.
3：List of Experimental Equipment and Materials:
A custom-made flow cytometer (LT-FCM) including a modulated laser source, novel single photon signal processing lifetime analysis electronics, and signal analysis and instrument control software was designed.
4：Experimental Procedures and Operational Workflow:
The interaction time of the encoded beads with the spot of the focused excitation laser was varied by electronically regulating the sheath fluid pressure.
5：Data Analysis Methods:
Luminescence lifetimes were calculated from intensity decay curves by means of a well-known equation, and numerical simulations based on synthetic decay curves were performed to study the impact of measurement parameters and conditions.
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