Protocol to isolate live single cells while retaining spatial information by combining cell photolabeling and FACS

Area of Science:

• Cell Biology
• Molecular Biology
• Cancer Research

Background:

• Single-cell analysis techniques are crucial for biological research.
• Current methods often result in the loss of spatial information from cells.
• Understanding cellular micro-environments is vital in fields like cancer research.

Purpose of the Study:

• To introduce a novel method for isolating live single cells from primary tumors.
• To preserve the spatial information of cells during the isolation process.
• To enable downstream single-cell omics analyses with retained spatial context.

Main Methods:

• Development of the photoconversion of areas to dissect micro-environments (PADME) approach.
• Integration of cell photolabeling with fluorescence-activated cell sorting (FACS).
• Isolation of live single cells from primary breast tumor samples.

Main Results:

• Successful isolation of live single cells while maintaining their spatial localization.
• Demonstration of the PADME method's efficacy in preserving cellular micro-environments.
• Generated single cells are viable for diverse downstream applications.

Conclusions:

• The PADME technique offers a powerful solution for spatial single-cell isolation.
• This method enhances the capabilities of single-cell omics studies, particularly in complex tissues.
• PADME facilitates deeper insights into tumor heterogeneity and cellular interactions.