Fluid-Squid: DIY Multiplexed Imaging of Cells and Tissues

Area of Science:

• Cell Biology
• Biotechnology
• Microscopy

Background:

• Multiplexed single-cell characterization offers deep biological insights but is often hindered by high costs and limited customizability.
• Existing technologies struggle to balance affordability with the need for tailored experimental workflows in cell biology.

Purpose of the Study:

• To develop Fluid-Squid, an open-source, cost-effective, and customizable automated fluidics platform for multiplexed imaging.
• To demonstrate the platform's capability in characterizing both single cells and tissue samples with high-plex antibody panels.
• To introduce novel barcoding strategies for pooled sample analysis, reducing experimental time, cost, and batch effects.

Main Methods:

• Development of Fluid-Squid, an integrated automated fluidics and microscopy system.
• Application of oligonucleotide-barcoded antibody panels for high-plex imaging (36-plex and 39-plex).
• Implementation of sample pooling and barcoding strategies for simultaneous multi-sample analysis.

Main Results:

• Successful imaging of human intestinal tissues and characterization of immune cells (splenocytes, PBMCs) using Fluid-Squid.
• Demonstration of lyophilization feasibility for multiplexed antibody panels.
• Validation of pooled sample analysis for antibody titration and blood preparation method assessment.

Conclusions:

• Fluid-Squid provides a flexible, cost-effective, open-source framework for advanced multiplexed imaging.
• The platform empowers researchers with adaptable solutions for single-cell and tissue characterization.
• This work significantly lowers barriers to entry for sophisticated multiplexed imaging experiments.