Courses
Protocols
- Whole Blood Intracellular Cytokine Staining (ICS) Protocol. This optimized whole-blood intracellular cytokine staining (ICS) protocol enables high-throughput detection of antigen-specific T-cell responses using a 96-well format. By stimulating fresh heparinized blood with mycobacterial antigens and applying standardized steps for red blood cell lysis, viability assessment, extracellular and intracellular staining, and flow-cytometric analysis, the method provides reliable immune-profiling data for vaccine evaluation, tuberculosis research, and HIV–TB immunology. Designed for efficiency and reproducibility, this ICS workflow supports advanced immunomonitoring in clinical and translational studies.
- 96-Well Flow Cytometry Protocol for Tissue-Derived Immune Cells. This 96-well plate flow cytometry protocol provides a standardized workflow for processing finely minced tissues, generating high-quality single-cell suspensions, and assessing antigen-specific immune responses through ELISA and multiparametric flow cytometry. The method includes enzymatic digestion with collagenases, mechanical dissociation, red blood cell lysis, viability assessment, and filtration to ensure optimal cell recovery and integrity. Cells are stimulated with mycobacterial antigens (PPD, TB10.4, H1) under defined conditions, followed by Fc blocking, dual-mode viability staining, extracellular and intracellular antibody labeling, and fixation for acquisition. This optimized, high-throughput protocol supports reliable immunomonitoring for studies in vaccinology, tuberculosis research, and translational immunology.
Biosafety