Serial flow cytometry in an inertial focusing optofluidic microchip for direct assessment of measurement variations
Matthew DiSalvo, Paul Patrone, Anthony J. Kearsley, Gregory A. Cooksey
Flow cytometry is an invaluable technology in biomedical research, yet it has limited ability to separate inherent sample variability from measurement uncertainty. This limitation makes it difficult to classify sample composition, find rare events, and compare measurements. Here we report a serial flow cytometer that measured each particle more than once along a flow path for direct uncertainty quantification (UQ). The optofluidic cytometer controlled particle positioning and velocities using a novel hybrid inertial and hydrodynamic particle focusing strategy and was automated for throughputs above 100 event s-1 with typical analysis yields better than 99.9 %. The real-time UQ provided by the serial cytometer identified flow conditions with fluorescence area measurement precision on the order of 1 %, as well as a range of well-tolerated low-sheath fluid flow conditions for increased sample throughput. The serial cytometer is anticipated to improve single-cell measurements through estimation (and subsequent control) of uncertainty contributions from various other instrument parameters leading to overall improvements in the ability to better classify sample composition and to find rare events.
, Patrone, P.
, Kearsley, A.
and Cooksey, G.
Serial flow cytometry in an inertial focusing optofluidic microchip for direct assessment of measurement variations, Lab on A Chip, [online], https://doi.org/10.1039/D1LC01169C , https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=933706
(Accessed December 9, 2023)