Overview

This article presents a protocol for subtractive patterning of live cell monolayers using a microfluidic probe (MFP). The method allows for local and selective lysis of adherent cells, facilitating downstream molecular profiling studies.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Microfluidics

Background

Cell patterning is essential for studying cell-to-cell interactions.
Traditional methods often involve lengthy protocols.
The MFP technology offers a rapid and efficient alternative.
Optimization of MFP parameters is crucial for stable experiments.

Purpose of Study

To develop a method for generating spatially defined cell patterns.
To enable real-time modifications of cell patterns.
To facilitate in-situ spatiotemporal analysis of cellular interactions.

Methods Used

Utilization of a microfluidic probe for local lysis of cells.
Preparation of cell monolayers on chamber slides.
Optimization of MFP parameters for hydrodynamic flow confinement.
Visual demonstrations to illustrate the procedure.

Main Results

Successful generation of defined cell patterns using MFP.
Real-time modification of patterns demonstrated.
Efficient local lysis of cells achieved.
Protocol allows for downstream analysis of cell lysates.

Conclusions

The MFP-based patterning technique is rapid and effective.
It provides a valuable tool for studying cell interactions.
Future applications may enhance understanding of cellular dynamics.

Frequently Asked Questions

What is the main advantage of using MFP technology?

The main advantage is the nearly instantaneous patterning capability, allowing for real-time modifications.

How are the cell monolayers prepared?

Cells are cultured in T-flasks, trypsinized, and seeded onto chamber slides.

What is the purpose of the cell-tracker dye?

The dye is used for visualization of cells during the patterning process.

What is hydrodynamic flow confinement?

It is a technique that localizes liquids using simultaneous injection and aspiration.

How does the MFP ensure coplanarity with the substrate?

Coplanarity is achieved by adjusting the tilt of the probe head using a goniometer.

Institution - IBM Research - Zurich

Articles Published in JoVE