Prof. douaa m. sayed validation and verification of flowcytometry areas

1. Validation and
Verification of
Flowcytometry
Douaa mohammed sayed
Director of Flow cytometry Lab
South Egypt Cancer Institute – Assiut University
DOUAA_SAYED2HOTMAIL.COM
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

5. Verification and Validation
Wikipedia definition
Verification and Validation, in engineering or quality management systems, it is the act of
reviewing, inspecting or testing, in order to establish and document that a product, service or
system meets regulatory or technical standards.
Validation – “Are you building the right thing?”
Verification – “Are you building it right?”

6. Quality Assurance in FCM
Why should we care?
Assurance that flow cytometry results you are generating are
due to the properties of the sample and not changes in:
●
• Instrument
●
• Assay performance
●
• Reagents
●
• Operator variation
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

7. INSTRUMENT
Basic Knowledge about the configuration of the machine is essential
It is important to know basic start up and shut down procedures along with routine
maintenance processes
MAINTENANCE log has to be maintained
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

8. INSTRUMENT
What all is to be monitored
◦ Optics
◦ Fluidics
◦ Electronics
How do you monitor
◦ Manual methods
◦ Automated methods
QC beads are very useful for monitoring and evaluating instrument performance
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

9. Flow Cytometry Beads
Spherotech Beads
Cytometer Setup and Tracking (CS&T) beads
Seven colour setup beads
Calibrite Beads
Flow-Check™ Fluorospheres
IMMUNO-BRITE™ Standards Kit
Flow-Check™ Pro Fluorospheres
Cell Counting Beads
Antibody Capture beads
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

10. Key Performance Factors in High
Quality Flow Cytometry Data
Relative measured values of fluorescence
Linearity and accuracy
Resolution of subpopulations, including dim subpopulations
Sensitivity
Reproducibility of results and cytometer performance
Tracking
Comparison of results across time and among laboratories
Standardization
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

11. Flow Cytometry Assay
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

12. Flowcytometry Assay
Verification
Confirm specifications
established by the manufacturer
Accuracy
Precision
Reportable range of results
Verify of manufacturer’s normal
range
Validation
In-house, home brewed
assay
• Accuracy
• Precision
• Analytical sensitivity
• Analytical specificity
• Reportable range
• Reference range
• Calibration procedures
• Control procedures
• Sample stability
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

13. Verification and Validation
Lot to Lot Antibody VERIFICATION
Setting up a new permeabilisation and fixing procedure using home
brewed reagents - Assay validation
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

55. Click icon to add table
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

80. ISO 15189
The
international standard
for medical
laboratory
quality and
competence
Seeking for accreditation of Flow cytometry laboratory
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

81. The ISO 15189 standard, designed specifically for medical laboratories, covers 15
management requirements and 8 technical requirements. The standard is
concise and to the point, with each section consisting of anything from a few
sentences up to several pages of explanation.
Management requirements Technical requirements
Organization and management
Quality management system
Document control
Review of contracts
Examination by referral laboratories
External services and supplies
Advisory services
Resolution of complaints
Identification and control of
nonconformities
Corrective action
Preventive action
Continual improvement
Quality and technical records
Internal audits
Management review
Personnel
Accommodation and environmental
conditions
Laboratory equipment
Pre-examination procedures
Examination procedures
Assuring quality of examination procedures
Post-examination procedures
Reporting of results
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

82. First Accredited Flow Cytometry Lab
Location: South Egypt Cancer
Institute – Assiut University
http://www.seci.info/
flowcytometry/
1st Arab Flow cytometry working group workshop 14th – 15th October 2016

88. 1st Arab Flow cytometry working group workshop 14th – 15th October 2016
Acknowledgements

89. Thank you
1st Arab Flow cytometry working group workshop 14th – 15th October 2016