Making deep immunophenotyping accessible: the successful application of a guided 23-parameter mouse immunophenotyping...

Area of Science:

• Immunology
• Biotechnology
• Analytical Chemistry

Background:

• High-parameter spectral flow cytometry enables detailed cellular analysis.
• Shared resource facilities (SRFs) can enhance researcher support through specialized panels.
• Developing standardized, accessible protocols is crucial for broad adoption.

Purpose of the Study:

• To develop and present a versatile 23-color mouse immunophenotyping panel.
• To provide a reproducible and cost-effective method for high-parameter spectral flow cytometry.
• To reduce barriers for researchers accessing advanced flow cytometry techniques.

Main Methods:

• Design and validation of a 23-color immunophenotyping panel for mouse samples.
• Optimization of reagents, staining protocols, and instrument setup.
• Development of acquisition and analysis templates, including manual gating and dimensionality reduction (tSNE, uMAP).

Main Results:

• The panel is broadly applicable across multiple tissue types.
• The protocol is modifiable, reproducible, and cost-effective.
• Implementation in a shared resource facility requires minimal staff preparation for maximum researcher benefit.

Conclusions:

• Shared resource facilities can offer significant value by providing accessible, high-parameter flow cytometry resources.
• This panel serves as a complete resource for institutions to adopt and adapt.
• The approach lowers entry barriers, increases reproducibility, and reduces costs for researchers in spectral flow cytometry.