Flow Cytometry
Flow Cytometry in Pediatric Diagnostics
Flow cytometry is a sophisticated laser-based technology that enables simultaneous analysis of multiple cellular parameters at the single-cell level. This technique is fundamental in pediatric diagnostics, particularly in hematology-oncology and immunology.
Key Points:
- Multiparameter cellular analysis
- Rapid analysis of thousands of cells
- Quantitative and qualitative assessment
- Essential for immunophenotyping
- Critical in diagnosis and monitoring of various conditions
Basic Principles:
- Forward scatter (cell size)
- Side scatter (cellular complexity)
- Fluorescence detection
- Immunophenotyping markers
- DNA content analysis
Clinical Applications in Pediatrics
Primary Diagnostic Applications:
- Hematologic Malignancies
- Acute leukemias classification
- Lymphoma diagnosis
- Minimal residual disease detection
- Disease progression monitoring
- Primary Immunodeficiencies
- SCID screening
- T-cell deficiencies
- B-cell deficiencies
- NK cell abnormalities
Monitoring Applications:
- Post-transplant monitoring
- Immunotherapy response
- Chimerism analysis
- Immune reconstitution
- Treatment effectiveness
Sample Collection and Processing
Collection Requirements:
- Specimen Types
- Peripheral blood
- Bone marrow
- CSF
- Tissue specimens
- Body fluids
- Collection Tubes
- EDTA for routine analysis
- Heparin for functional studies
- Special media for specific tests
Critical Factors:
- Time constraints
- Process within 24-48 hours
- Temperature control
- Light protection
- Sample viability
- Cell count requirements
- Viability assessment
- Quality control measures
Analysis Parameters and Markers
Common Marker Panels:
- T-Cell Markers
- CD3, CD4, CD8
- CD45RA, CD45RO
- TCRαβ, TCRγδ
- B-Cell Markers
- CD19, CD20
- CD10, CD34
- Surface Immunoglobulins
- Myeloid Markers
- CD13, CD33
- CD14, CD64
- CD117, HLA-DR
Special Applications:
- Intracellular Markers
- Cytokine Analysis
- Cell Cycle Analysis
- Functional Studies
Result Interpretation and Reporting
Interpretation Guidelines:
- Quality Control Metrics
- Internal controls
- Viability assessment
- Background fluorescence
- Compensation settings
- Analysis Strategies
- Gating hierarchies
- Population identification
- Quantitative assessment
- Pattern recognition
Reporting Elements:
- Population percentages
- Absolute counts
- Marker expression patterns
- Reference ranges
- Clinical correlation
Disease-Specific Analysis Patterns
Leukemia/Lymphoma:
- ALL Classification
- B-ALL markers
- T-ALL markers
- Aberrant expression
- MRD assessment
- AML Analysis
- Lineage assessment
- Maturation patterns
- Aberrant phenotypes
Immunodeficiencies:
- SCID Screening
- T-cell enumeration
- Naive T-cell assessment
- B and NK cell quantification
- Other PIDs
- Specific marker deficiencies
- Functional assessments
- Memory cell evaluation
