In Vitro Photoacoustic Flow Cytometry: A Non-invasive Flow Photoacoustic Technique to Detect Nanoparticle-bearing Circulating Ovarian Cancer Cells

Abstract

Source: Joel F. Lusk et al. Ovarian Cancer Detection Using Photoacoustic Flow Cytometry. J. Vis. Exp. (2020)

In this video, we present a photoacoustic flow cytometric detection of ovarian cancer cells bearing nanoparticles in a circulating in vitro flow chamber. This non-invasive technique successfully identifies circulating tumor cells in a flow system utilizing a contrast agent.

Protocol

Flow System Architecture 1. Flow chamber construction Using the provided Solid Works file, 3D print the flow tank using ABS thermoplastic or PLA plastic. Dimensions are provided below if SolidWorks is unavailable. Figure 1A shows a representation of this model. The body of the flow tank is 2.5 cm x 1.5 cm x 7.5 cm. The far ends of the flow tank include holes approximately 5 mm in diameter to allow for the entry of tubing co…

Representative Results

Figure 1. Representative images of photoacoustic flow cytometry system and flow chamber. (A) Detailed view of the 3D printed flow chamber. (B) Diagram of PAFC system. (C) Flow system architecture: SP = syringe pump; DAQ/FPGA = data acquisition/field programmable gate array; Ob = objective len…

Materials

3D Printed Tank		Custom-made
Acquisition Card	National Instruments	PXIe-5170R	250 MS/s, 8-Channel, 14-bit
Alconox	Sigma-Aldrich	242985-1.8KG	Detergent used for cleaning glassware.
Amicon Ultra-15 Centrifugal Filters	Millipore	UFC903024
Amicon Ultra-4 Centrifugal Filters	Millipore	UFC803024
Data Acquisition software	National Instruments	NI LabVIEW 2017 (32-bit)	LabVIEW used to synchronize laser pulses with data acquisition.
Data Processing Software	Mathworks	Matlab R2016a	Reconstructions and graphs produced using Matlab software.
FBS	Sigma-Aldrich	F2442-500ML
Pulsed Laser	RPMC Lasers Inc	Quantus-Q1D-1053	Pulsed laser source with specifications 1053 nm, 8 ns pulse, 10 Hz maximum.
Pulser/Receiver	Olympus	5077PR	Receives, filters, and amplifies photoacoustic signals. Operated with 59 dB Gain.
Quartz Capillary Tube	Sutter Instrument	QF150-75-10
Syringe Pumps	New Era Pump Systems Inc	DUAL-1000
Coupling Stage	Newport	F-91-C1-T	Stage for coupling pulsed light to objective. Holds FP-1A and LMH-10x-532
Coupling Objective	Thorlabs	LMH-10x-532	To couple pulsed light to optical fiber.
Norm-Ject 10 mL Syringes	HENKE SASS WOLF	4100-X00V0
Transducer	Olynmpus	V214-BB-RM	Ultrasound detector with central frequency of 50 MHz and -6 dB fractional bandwidth of 82%.
Trypan Blue Solution 0.4%	Amresco	K940-100ML
Optical Fiber	Thorlabs	FG550LEC	Used to expose sample to pulsed light.
Tween 20	Sigma-Aldrich	P7949-100ML