G. slide storage – Leica Biosystems HER2 FISH System - 30 Test User Manual

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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 first 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. Confirm 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 field displays Proc (OK) and batch number and finish 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 fluorescence

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.

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