Intracellular Flow Cytometry Improvements in Clinical Studies

Area of Science:

• Immunology and Cell Biology
• Clinical Diagnostics

Background:

• Intracellular flow cytometry is vital but faces challenges in routine clinical use and multicenter studies.
• Current methods are time-consuming, cause cell loss, and lead to nonspecific binding or reduced fluorescence, hindering standardization.

Purpose of the Study:

• To present novel, clinic-ready intracellular flow cytometry protocols.
• To overcome limitations of existing methods for improved standardization and feasibility in clinical settings.

Main Methods:

• Development of one-step, whole-blood, lyse-no-wash-no-centrifuge protocols.
• Utilization of improved staining quality and lyophilized reagents in ready-to-use tubes.
• Application in immunomonitoring of septic patients, evaluating myeloperoxidase, lactoferrin, FOXP3, and STAT5 phosphorylation.

Main Results:

• Protocols offer shorter time-to-results and improved staining quality.
• One-step procedures minimize cell loss and nonspecific binding.
• Demonstrated feasibility in evaluating specific biomarkers in neutrophils and lymphocytes.

Conclusions:

• Novel protocols enhance standardization and feasibility of intracellular flow cytometry in clinical studies.
• These advancements support drug efficacy monitoring and patient stratification for personalized medicine.
• The methods are applicable to immunomonitoring in conditions like sepsis.