A flow cytometer that uses sound waves to determine the size of biological cells is presented. In this system, a microfluidic device made of polydimethylsiloxane (PDMS) was developed to hydrodynamically flow focus cells in a single file through a target area. Integrated into the microfluidic device was an ultrasound transducer with a 375 MHz center frequency, aligned opposite the transducer was a pulsed 532 nm laser focused into the device by a 10x objective. Each passing cell was insonfied with a high frequency ultrasound pulse, and irradiated with the laser. The resulting ultrasound and photoacoustic waves from each cell were analyzed using signal processing methods, where features in the power spectra were compared to theoretical models to calculate the cell size. Two cell lines with different size distributions were used to test the system: acute myeloid leukemia cells (AML) and melanoma cells. Over 200 cells were measured using this system. The average calculated diameter of the AML cells was 10.4 ± 2.5 μm using ultrasound, and 11.4 ± 2.3 μm using photoacoustics. The average diameter of the melanoma cells was 16.2 ± 2.9 μm using ultrasound, and 18.9 ± 3.5 μm using photoacoustics. The cell sizes calculated using ultrasound and photoacoustic methods agreed with measurements using a Coulter Counter, where the AML cells were 9.8 ± 1.8 μm and the melanoma cells were 16.0 ± 2.5 μm. These results demonstrate a high speed method of assessing cell size using sound waves, which is an alternative method to traditional flow cytometry techniques.
Eric M. Strohm, Vaskar Gnyawali, Mia Van De Vondervoort, Yasaman Daghighi, Scott S. H. Tsai, and Michael C. Kolios, "Classification of biological cells using a sound wave based flow cytometer," Proc. SPIE 9708, Photons Plus Ultrasound: Imaging and Sensing 2016, 97081A (Presented at SPIE BiOS: February 15, 2016; Published: 15 March 2016); https://doi.org/10.1117/12.2211740.
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