Multi-color flow cytometer with PIC-based structured illumination.

Area of Science:

• Biophotonics
• Optical Engineering
• Analytical Chemistry

Background:

• Traditional flow cytometry struggles to definitively assign multicolor signals to specific excitation sources.
• Accurate signal attribution is crucial for multiplexed analysis and reducing spectral overlap.

Purpose of the Study:

• To demonstrate a novel flow cytometer design utilizing structured light illumination.
• To enable precise attribution of fluorescent and scattering signals to their excitation wavelengths.

Main Methods:

• Development of a multicolor light source with structured illumination patterns (405, 488, 561, 640 nm) using a silicon nitride photonic integrated circuit.
• Conducting cytometry experiments with calibration beads.
• Comparing performance metrics against a commercial flow cytometer.

Main Results:

• The novel flow cytometer successfully attributes signals to excitation wavelengths using structured illumination.
• The experimental device showed comparable performance to a commercial device, with a slightly higher sensitivity floor.
• Most stained beads were accurately categorized, validating the concept's feasibility.

Conclusions:

• Structured light illumination is a feasible and effective method for enhancing multicolor flow cytometry.
• This approach offers improved spectral attribution and potential for more accurate multiplexed analysis.