HercepTest™ Interpretation Manual - Breast - Dako

Troubleshooting Guide
Troubleshooting Guideline for HercepTest TM
Problem
Probable Cause
Suggested Action
Reference
1. No staining
of slides
2. Weak staining
of slides
1a. Programming error. Reagents Check programming grid to verify that the
not used in proper order. staining run was programmed correctly.
1b. Reagent vials were not loaded Check the Reagent Map to verify the proper
in the correct locations in the location of reagent vials.
reagent racks.
1c. Insufficient reagent on Ensure that enough reagent is loaded into
tissue section.
the reagent vials prior to commencing the
run. Refer to the Reagent Map for volumes
required. Ensure that spreading of reagent
is optimal.
1d. Sodium azide in
Use fresh preparation of Wash Buffer
Wash Solution.
provided in the kit.
1e. Excessive heating for more Air dry the tissue sections at room
than one hour at ≥ 60 °C temperature for a minimum of 12 hours
may cause a significant or until dry. Alternatively, dry at 37 °C
decrease or loss of the overnight or dry at 60 °C for a maximum
specific membrane-associated of one hour. Drying of tissue sections at
HER2 immunoreactivity. elevated temperatures must only be
performed in a calibrated oven with
uniform heat distribution.
2a. Inadequate epitope retrieval. Verify that Epitope Retrieval Solution
reaches 95-99 °C for full 40 minutes and is
allowed to cool for an additional 20 minutes.
HercepTest TM Interpretation
Manual Artificial
Heterogeneous Staining
(page 20)
HercepTest TM Interpretation
Manual Excessive Tissue
Drying. Loss of specific
staining (page 30)
HercepTest TM Interpretation
Manual Effects of
Insufficient Target
Retrieval. (page 29)
2b. Inadequate reagent
incubation times.
2c. Inappropriate fixation
method used.
2d. Excessive heating for more
than one hour at ≥ 60 °C
may cause a significant
decrease or loss of the specific
membrane-associated
HER2 immunoreactivity.
Review Staining Procedure instructions.
Ensure that patient tissue is not over-fixed
or that an alternative fixative was not used.
Air dry the tissue sections at room
temperature for a minimum of 12 hours or
until dry. Alternatively, dry at 37 °C overnight
or at 60 °C for a maximum of one hour.
Drying of tissue sections at elevated
temperatures must only be performed
in a calibrated oven with uniform heat
distribution.
See
Instructions for Use
HercepTest TM Interpretation
Manual Effects of Fixation.
(page 28)
HercepTest TM Interpretation
Manual Effects of
Excessive Tissue Drying.
Loss of specific staining
(page 30)
46
HercepTest TM Interpretation Manual – Breast Cancer
ROW Version