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Leica Biosystems Leica HER2 FISH System - 30 Test Instructions for Use TA9217 EN-CE-Rev_D 08/04/2013
5. Turn on the PC attached to the Leica BOND-MAX and BOND-III System.
6. Open the Leica BOND software.
7. For a new Leica HER2 FISH System - 30 Test kit, scan the reagent tray barcode with
the handheld
scanner to enter the system into the Leica BOND reagent inventory (Single barcode
only).
8. Prepare Leica BOND Enzyme 5 in the supplied Leica BOND Open Container at a dilution
of 1:300. For example, for 10 slides add 10µL of Leica BOND Enzyme Concentrate 2 to
2990 µL of Leica BOND Enzyme Diluent.
9. Scan in supplied Leica BOND Open Container and register as Bond Enzyme 5.
10. Go to the Slide setup screen and click Add case.
11. Enter details for the rst case. Ensure the dispense volume is set to 150 µL and the
preparation protocol is *Dewax. Click OK.
12. With the case highlighted in the Slide setup screen click Add slide.
13. First, add patient test slides. Ensure tissue type is set to Test tissue.
14. Select staining mode Single.
15. Select process ISH.
16. Select *LSI HER2/CEP17 Dual Probe – 30 Test from the probe list. The Protocols tab
defaults to the correct staining protocol (*FISH Protocol A), HIER protocol (*HIER 25
min with ER1 (97)), EIER protocol (*Enzyme 5 for 25 min), denaturation (*D10) and
hybridization (*ISH Hybridization (12Hr)).
17. Repeat steps 10 to 16 until patient test slides and controls (Leica HER2 FISH control
slides and/or in-house controls) have been created. Print slide labels.
18. Label slides appropriately.
19. Open the lids of all Leica HER2 FISH System - 30 Test containers and load the reagent
tray onto the Leica BOND-MAX and BOND-III System.
20. Apply new Covertiles to each slide.
21. Load the slide tray onto the Leica BOND-MAX and BOND-III System and press the
Load/Unload button.
22. Conrm that the slides have been scanned and click the Run (Play) button on the System
status screen to commence the run immediately (for the Leica HER2 FISH System -
30 Test it is recommended that this assay is run overnight utilizing the delayed start
functionality).
23. Ensure that the tray indicator eld displays Proc (OK) and batch number and nish time
are displayed.
24. When the run is completed press the Load/Unload button and remove the slide tray from
the Leica BOND-MAX and BOND-III System.
25. Remove Covertiles and rinse the slides in de-ionized water.
26. Dehydrate rapidly in two changes of alcohol, air dry.
27. Dispense 20µL of DAPI directly onto the sample.
28. Apply coverslip and allow the solution to spread to its full extent, taking care to remove
any air bubbles.
29. Seal edge of coverslip with nail varnish, or similar sealant.
30. Place slides in dark to facilitate signal development before viewing under uorescence
microscope.
31. To preserve signal intensity store stained slides at -20 °C.
G. Slide Storage
Store stained slides at -20 °C in the dark. Allow slides to reach room temperature prior to
viewing following removal from -20 °C.