In Situ Hybridization - Cost and Assurances

 

No guarantees can be made for the successful application of in situ hybridization histochemistry using new oligonucleotide probes. However several assurances can be given based on experience and use of appropriate controls along the way. The procedure is divided into steps which are only performed following the successful completion of the prior step. Investigators will be responsible for the cost of any step performed except when we demonstrate that the problem was related to us. Since we always include a labeling and hybridization control, we are able to identify those problems. All costs associated with the preliminary steps of probe design, probe purchase, probe preparation and/or consultation will have to be paid when completed. In addition, all tissue related costs for subbing slides, sectioning of tissue and tissue storage would also be paid when completed. Investigators will be responsible for payment of labeling of their probe. At the time of each labeling, we also label one of our housekeeping control oligonucleotide probes. In the event that the labeling does not reach 50% incorporation in our housekeeping probe, the investigator will not be responsible for the labeling cost of that reaction. However, if we successfully label 'our' housekeeping probe, the investigator will be responsible for our attempt to label 'their' probe(s) of interest. Failure to adequately label a probe can be due to defective synthesis of the probe, degradation of the probe, or contamination which we cannot control. In the event of a failed labeling, investigators will be offered the choice of either another attempt at labeling, going ahead with a marginally labeled probe, or diluting or remaking their oligonucleotide probe.
If both the investigators' probe of interest and the housekeeping probe are successfully labeled we will proceed with tissue hybridization. During hybridization we will use our housekeeping probe with 'your' tissue and with 'our' tissue. In the event that our labeled housekeeping probe does not properly hybridize with 'our' tissue the investigator will not be responsible for payment of that hybridization portion of the procedure. In all other cases, the investigator will be responsible for the cost of the procedure whether or not 'their' probe properly labels 'their' tissue. The failure to achieve proper hybridization can be due to degradation of mRNA within 'your' tissue, improper probe design or very low abundance and we cannot be responsible for those issues. We will also hybridize our control housekeeping probe with your tissue to determine mRNA quality and abundance that might guide subsequent steps.
Investigators will have the ability to review the first hybridization and discuss it. Depending on the outcome, the investigator might choose to do another round of test labeling and hybridization or proceed directly to hybridization in their experimental tissue. A second test procedure is usually recommended to optimize and verify the hybridization and post wash temperatures. It is highly recommended prior to performing an experiment using a large number of samples. Similar control steps will be taken with each labeling and hybridization.
Investigators will also be responsible for all post hybridization costs which might include densitometry and image capture, emulsion dipping, development and staining, and/or grain counting or image capture.

 

Molecular Histology Center Environmental and Occupational Health Sciences Institute, 170 Frelinghuysen Road, Piscataway, NJ 08854 Phone: 732-445-3729 For additional information contact ekr@eohsi.rutgers.edu

Updated on Friday, June 03, 2005