Validation, verification, or optimization? Don’t guess – here’s how to get it right!
A pathologist just asked you to bring in a new antibody or special stain – now what? Maybe you’ve got a fresh control block that needs approval, or you’re swapping products to a new vendor. Should you do a validation or verification? Or maybe an optimization? What’s the right approach? Let’s break it down in a way that makes sense.
Optimization: Finding the Sweet Spot
Optimization is a trial-and-error phase – tweaking protocols to get the best staining results. This helps fine-tune specificity and signal strength.
When do you optimize?
- Introducing a new antibody or clone
- Working with new H&E or special stains
- Adjusting preanalytical factors (processing schedules, fixation times, etc.)
- When a pathologist requests changes to a stain or antibody
Mart 1 optimized in two different labs
Validation: The Gold Standard of Confidence
Think of validation as the ultimate test drive. It confirms that a test or stain is specific, accurate, and reproducible – giving you confidence that every time you run it, you get reliable results. Before using any test on patient samples, it must be validated.
When do you validate?
Absolutely! CAP and CLIA set the rules and recommendations, especially for antibody validation. Here are some of the standards and guidelines to look at:
Think of validation as the ultimate test drive. It confirms that a test or stain is specific, accurate, and reproducible – giving you confidence that every time you run it, you get reliable results. Before using any test on patient samples, it must be validated.
When do you validate?
- Introducing a new test
- New antibody or new clone of established antibody
- Different fixative
- New detection kit
- Decalcification
- New platform
Absolutely! CAP and CLIA set the rules and recommendations, especially for antibody validation. Here are some of the standards and guidelines to look at:
- CAP: ANP.22750 Antibody Validation/Verification
- CAP: ANP.22978 Predictive Marker Testing – Validation/Verification
- CAP: ANP.22985 Predictive Marker Testing – Decalcified Specimens
- CLIA: § 493.1253 Standard - Establishment of performance specifications
S100 Monoclonal
S100 Polyclonal
Verification: The Quality Check
Already have a validated test but need to make a tweak? That’s where verification comes in. It ensures that any changes still meet staining quality expectations.
When do you verify?
Yes! Just like with validation, CAP and CLIA set the rules and recommendations for verification. Here are some of the standards and guidelines to look at:
Already have a validated test but need to make a tweak? That’s where verification comes in. It ensures that any changes still meet staining quality expectations.
When do you verify?
- Switching manufacturers of an established clone
- Changing antigen retrieval methods
- Platform is moved
- Minor protocol changes (antibody dilution, incubation times)
- Checking antibody lot to lot
- Test control blocks
- Using FDA-approved kits
Yes! Just like with validation, CAP and CLIA set the rules and recommendations for verification. Here are some of the standards and guidelines to look at:
- CAP: ANP.22750 Antibody Validation/Verification
- CAP: ANP.22978 Predictive Marker Testing – Validation/Verification
- CAP: ANP.22780 ISH Assay Performance
- CLIA: § 493.1253 Standard - Establishment of performance specifications
Mart 1 Clone A103 from three different manufacturers
So, What Do You Need?
Be sure to document everything to show evidence of compliance:
- Validation → New test, major changes, or new platforms
- Verification → Adjustments to an already validated test
- Optimization → Fine-tuning for the best possible results
Be sure to document everything to show evidence of compliance:
- SOP: acts as your roadmap for validation, verification, or optimization process
- Number of positive and negative tissues required
- Any exemptions
- Route to approval
- Protocols used
- Records
- How or where tissue was sourced
- Lot numbers, clones, expiration dates, manufacturer, or any other information
- Run reports
- Record or form of approval that is signed
- Slides/Results
- Maintain for a minimum of two years (check your SOPs)
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