Rapid one-step protocol for exploring immune cells in cerebrospinal fluid

Area of Science:

• Neuroimmunology
• Clinical Diagnostics
• Flow Cytometry

Background:

• Cerebrospinal fluid (CSF) immune cell profiling aids in diagnosing nervous system pathologies.
• Current methods face challenges due to rapid cell degradation, limited sample volume, and non-standardized staining protocols.
• There is a critical need for improved pre-analytical and analytical processes for CSF immune cell analysis.

Purpose of the Study:

• To develop and validate a one-step flow cytometry protocol for analyzing CSF immune cells.
• To assess the repeatability, stability, and impact of sample processing delays on CSF immune cell analysis.
• To establish a rapid, standardized, and reproducible method for multiparametric CSF immune cell analysis.

Main Methods:

• Developed a one-step antibody-based protocol for CSF cell analysis without a washing step.
• Validated the protocol's repeatability and stability over 3 days using mild fixation.
• Assessed the impact of post-collection delays on cell counts and immune staining.

Main Results:

• The one-step protocol demonstrated high repeatability and stability for immune cell subset counts and staining over 3 days.
• Direct labeling without washing, coupled with cytometer performance, ensures precise and reproducible cell counting.
• Significant loss of monocytes was observed in samples not processed rapidly after lumbar puncture.

Conclusions:

• The developed one-step protocol provides a rapid, standardized, and reproducible method for multiparametric CSF immune cell analysis.
• This protocol addresses limitations of existing methods, improving diagnostic and prognostic capabilities for neurological conditions.
• Timely processing of CSF samples is essential to prevent the loss of critical immune cell populations like monocytes.