Leica Biosystems HER2 FISH System - 30 Test - Manual (Page 21)

Page 21 of 24
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Leica Biosystems Leica HER2 FISH System - 30 Test Instructions for Use TA9217 EN-CE-Rev_D 08/04/2013
English
Troubleshooting
Problem Probable Cause Remedial Action
No or weak
fluorescent
signal/staining Inappropriate fixation or
processing of test specimen Ensure a formalin-based fixative is used and
that processing schedules are suitable for
the specimen undergoing testing.
Leica HER2 FISH System - 30
Test is being used outside its
expiry date Ensure the Leica HER2 FISH System - 30
Test used is within its specified expiry date.
Incorrect protocol selection Ensure appropriate default to *FISH Protocol
A in the staining protocol field of the Add slide
dialog.
Inappropriate bulk reagents
dispensed Ensure all Leica BOND reagents have been
allocated to appropriate bulk containers
and placed into appropriate positions on the
instrument.
Inadequate deparaffinization
of slides Ensure *Dewax mode is selected in the
Preparation field of the Add slide dialog.
Inappropriate pretreatment Ensure default pretreatment (HIER and
Enzymatic Digestion) protocols are selected.
Adjust pretreatment protocol (HIER or
Enzymatic Digestion) if required.
Inadequate denaturation Ensure appropriate default denaturation *D10 is
selected.
Inadequate hybridization Ensure appropriate default hybridization *H12 is
selected. Extend hybridization time if required.
Excess post hybridization
washing Decrease post hybridization wash incubation
time.
Run aborted prior to
completion Using Leica BOND software, confirm the
presence of any reportable errors during the
staining run and address as instructed by
the Leica BOND software.
Inappropriate fluorescence
microscopy equipment
•
Inappropriate filter set
• Incorrect lamp
• Expired lamp
• Incorrect oil type
Ensure that all fluorescence microscopy
equipment used is appropriate for the assay
being performed, confirm:
•
Appropriate filter set
• Appropriate lamp
• Good lamp strength
• Appropriate oil for use in oil immersion
microscopy
Over exposure to UV light
(photobleaching) Store slides before and after assessment in
the dark to preserve fluorescent signals. To
maintain signal for long term storage store
slides at –20 ºC.