Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry.

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Flow cytometry is a cornerstone for analyzing hematopoietic cell phenotypes.
• Conventional flow cytometry faces challenges with multi-parametric analysis and auto-fluorescence.
• Increasing complexity of fluorescent probes and the need to analyze auto-fluorescent tissues necessitate advanced methods.

Purpose of the Study:

• To describe a spectral flow cytometry protocol for enhanced cell analysis.
• To demonstrate the capability of spectral flow cytometry in handling multi-parametric data and auto-fluorescence.
• To showcase the advantages of spectral flow cytometry for analyzing complex biological samples.

Main Methods:

• Utilized spectral flow cytometry for high-dimensional analysis.
• Employed an algorithm to replace compensation matrices and manage auto-fluorescence.
• Performed 21-parameter characterization using 19 fluorescent probes.

Main Results:

• Spectral flow cytometry effectively discriminates fluorochromes with similar emission peaks.
• The method provides multi-parametric analysis without compensation requirements.
• Successfully analyzed auto-fluorescent cell suspensions from challenging tissues like the heart and intestine.

Conclusions:

• Spectral flow cytometry offers superior analytical capabilities compared to conventional methods.
• This technique overcomes compensation matrix complexity and manages auto-fluorescence.
• Enables high-resolution detection of minor cell populations in difficult-to-analyze tissues.