LABORATORY OPERATIONS Flashcards
How should an optimal positive control be produced?
In the exact same conditions as the patient tissue (fixation, processing, staining) and cut onto the same slide.
How can you validate a prediluted antibody?
By performing a serial dilution (1:2, 1:4, 1:8 and 1:16) and assessing for reactivity on known positive tissue.
What is the one prediluted antibody that can be run without additional lot validation?
FDA approved pre diluted antibody
Define concentrated antibody
A non-diluted antibody in which several serial dilutions must be performed in order to find the optimal dilution. Called “prep kits” in Vancouver.
Are diluted or concentrated antibody more practical?
Concentrated
Much more shelf stable and are much cheaper than diluted antibody
Concentrated antibody also give the user much more control over antibody expression in patient tissue
How do you make a1:20 diluted antibody?
1 part antibody
19 parts diluent totaling up to 20 parts
What is a lambda in reference to measurement?
An old fashioned way to say microliter
Same as micron=micrometer
Define reagent control
Define procedure control
Define a positive tissue control
Define a negative tissue control
Define an internal tissue control
What are the four common negative control reagents used in place of the primary antibody?
1.
2.
3.
4.
What two negative control reagents detect non-specific background staining?
Which control types validate the staining steps?
Which control types validate fixation and processing?
Which antigen can be used as a control to evaluate fixation quality?
Vimentin
Which control type assists with evaluation of prognostic and predictive IHC assays?
What are the three characteristics of an optimal positive control?
1.
2.
3.
What are the optimal storage options for control tissue?
What are the advantages of purchased control tissue?
What are the disadvantages of purchased control tissue?
What issues must be considered when selecting control tissues?
Explain the use of multi-tissue blocks
Define lot validation
Describe a method for lot validation
Explain procedure validation
Describe a method for procedure validation
Explain antibody validation
Describe a method for antibody validation
Describe a method for control tissue validation
What records are recommended for control validations?
Define ASR
What regulations pertain to ASR?
Define IVD
What regulations pertain to IVD?
Define RUO
What regulations pertain to RUO?
How does the ASR/IVDF/RUO
