Home › Immunology › Detection and Isolation of Circulating Melanoma Cells using Photoacoustic Flowmetry
Immunology JoVE (Open Access) Citable · DOI

Detection and Isolation of Circulating Melanoma Cells using Photoacoustic Flowmetry

DOI: 10.3791/3559-v
What you'll learn
  • ✓ Prepare blood samples and construct a flow chamber for circulating cell detection
  • ✓ Set up and operate a photoacoustic flowmetry system using laser-induced ultrasound
  • ✓ Isolate and identify circulating melanoma cells via immunocytochemistry
  • ✓ Apply photoacoustic detection as a metastatic disease screening method
Protocol

We have developed a flow cytometer using laser induced ultrasound to detect circulating melanoma cells as an early indicator of metastatic disease.

Difficulty
advanced
Total time
~4–6 hours per sample (including blood preparation, chamber setup, flowmetry acquisition, and immunostaining)
Biosafety
BSL-2

Steps

1
Prepare blood sample for circulating cell analysis

Process whole blood or plasma samples to isolate circulating cells for downstream detection. Standard anticoagulation and handling protocols are applied to preserve cell viability.

▶ 01:20
2
Construct flow chamber for photoacoustic detection

Assemble a microfluidic or macro flow chamber with optical and acoustic access windows to enable cell transit monitoring and photoacoustic signal acquisition.

▶ 02:58
3
Set up photoacoustic flowmetry instrumentation

Configure the laser system, acoustic transducers, and detection electronics to enable real-time monitoring of circulating cells via laser-induced ultrasound signals as cells flow through the chamber.

▶ 04:32
4
Perform immunocytochemistry on isolated cells

Apply fluorescent antibody staining to phenotypically confirm melanoma cell identity and distinguish them from background blood cells in captured populations.

▶ 06:42
5
Analyze results and isolate melanoma cells

Review photoacoustic detection data and correlated immunofluorescence images to identify, enumerate, and characterize circulating melanoma cells as potential metastatic indicators.

▶ 08:39
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