Home › Biochemistry › Rapid Subtractive Patterning of Live Cell Layers with a Microfluidic Probe
Biochemistry JoVE (Open Access) Citable · DOI

Rapid Subtractive Patterning of Live Cell Layers with a Microfluidic Probe

DOI: 10.3791/54447-v
What you'll learn
  • ✓ Operate a microfluidic probe for selective cell lysis and patterning
  • ✓ Create hierarchical hydrodynamic flow confinement on live cell monolayers
  • ✓ Retrieve and process cell lysates for molecular profiling analysis
  • ✓ Design and execute subtractive patterning experiments on adherent cells
Protocol

We present a protocol to perform subtractive patterning of live cell monolayers on a surface. This is achieved by local and selective lysis of adherent cells using a microfluidic probe (MFP). The cell lysate retrieved from local regions can be used for downstream analysis, enabling molecular profiling studies.

Difficulty
advanced
Total time
~4–6 hours per experiment (including MFP setup, patterning, and downstream processing)
Biosafety
BSL-1

Steps

1
Clean and prepare microfluidic probe head and platform

Perform thorough cleaning and calibration of the MFP apparatus and ancillary equipment. Prepare the platform for selective cell lysis experiments.

▶ 01:16
2
Create hierarchical hydrodynamic flow confinement

Establish hierarchical HFC parameters and apply the microfluidic probe to initiate local, controlled flow confinement over the cell monolayer for selective lysis.

▶ 06:13
3
Process lysate samples and amplify extracted DNA

Collect retrieved cell lysate from patterned regions and perform downstream molecular processing, including DNA extraction and amplification for profiling studies.

▶ 08:10
4
Evaluate patterning results and data quality

Assess the efficacy of subtractive patterning on cell monolayers and validate the quality of molecular data obtained from lysate analysis.

▶ 09:46
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