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…
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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 |
