Single Cell Microarrays Fabricated by Microscale Plasma-Initiated Protein Patterning (μPIPP)

Area of Science:

• Cell Biology
• Biophysics
• Biotechnology

Background:

• Single cell fluorescence time-lapse measurements are crucial for understanding cellular dynamics.
• Standardized boundary conditions are essential for reproducible single cell assays.
• Existing methods for creating cell-patterned arrays can be complex or restrictive.

Purpose of the Study:

• To provide a detailed and reproducible protocol for fabricating high-quality single cell arrays.
• To demonstrate the utility of these arrays for various single cell measurements.
• To highlight the advantages of microscale plasma-initiated protein patterning (μPIPP) for array generation.

Main Methods:

• Microscale plasma-initiated protein patterning (μPIPP) for creating microstructured protein islands.
• Fabrication of nonadhesive areas to isolate individual cells.
• Automated data acquisition and analysis for time-lapse measurements.

Main Results:

• Successfully fabricated high-quality single cell arrays with reproducible results.
• Demonstrated efficient measurement of single cell gene expression after mRNA transfection.
• Showcased time courses of single cell apoptosis using the developed arrays.

Conclusions:

• μPIPP offers an easy and versatile method for fabricating single cell arrays on various substrates with diverse proteins.
• The developed arrays facilitate efficient, high-throughput analysis of single cell dynamics.
• This protocol supports the assessment of single cell dynamics using fluorescent reporters for broad applications in cell biology.