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

Published: April 30, 2023

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…

Açıklamalar

The authors have nothing to disclose.

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

Etiketler

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Bu Makaleden Alıntı Yapın
In Vitro Photoacoustic Flow Cytometry: A Non-invasive Flow Photoacoustic Technique to Detect Nanoparticle-bearing Circulating Ovarian Cancer Cells. J. Vis. Exp. (Pending Publication), e20369, doi: (2023).

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