Further Information; Date Of Issue - Leica BIOSYSTEMS BOND Hybridization Solution AR9037 Mode D'emploi

Instructions for Use
Dilute the ISH probe concentrate to the user-determined concentration with BOND Hybridization Solution. Transfer the diluted probe to a
BOND Open Container and register as an ISH probe on the BOND system (includes BOND-MAX system and BOND-III system).
Probe dilutions must be tested using the appropriate positive control tissue for the particular ISH probe (see "Quality Control" in the
"Using BOND Reagents" section of your user documentation).
When preparing dilutions for titration, ensure that allowance is made for the dead volume of the appropriate BOND Open Container (for
values refer to your BOND user documentation).
Product Specific Limitations
BOND Hybridization Solution was developed for use on the automated BOND system (includes BOND-MAX system and BOND-III
system).
The recommended pre-treatment and staining protocol are determined by the specific probe (and detection) selected. Please refer to the
relevant Instructions for Use for further information.
Users who deviate from recommended test procedures must accept responsibility for interpretation of patient results under these
circumstances. Protocol times may vary due to variation in tissue type, fixation and processing. In addition BOND Enzyme concentration
and incubation time may require optimization depending on tissue type, processing and fixation conditions.
Negative reagent controls should be used for CISH when optimizing pre-treatment conditions and protocol times.
The appropriate concentration of user's own ISH probe may vary, due to variation in tissue fixation and the effectiveness of enzymatic
digestion, and must be determined empirically.
Troubleshooting
Reference 3 may aid in remedial action.
Test samples should be complemented by the appropriate tissue and reagent controls.
Contact your local distributor or the regional office of Leica Biosystems to report unusual staining.

Further Information

Further information on ISH with BOND reagents, under the headings Principle of the Procedure, Materials Required, Specimen
Preparation, Quality Control, Assay Verification, Interpretation of Staining, Key to Symbols on Labels, and General Limitations can be
found in "Using BOND Reagents" in your BOND user documentation.
Bibliography
1. Clinical Laboratory Improvement Amendments of 1988, Final Rule 57 FR 7163 February 28, 1992.
2. Villanova PA. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory workers from infectious
diseases transmitted by blood and tissue; proposed guideline. 1991; 7(9). Order code M29-P.
3. Wilkinson DG. The theory and practice of in situ hybridization. In: Wilkinson DG. (ed.) In situ Hybridization. A practical approach. 2nd
Edition. New York: Oxford University Press, 1998, pp.18–20.

Date of Issue

03 October 2018
AR9037
Page 3