Protocols

This is a fee-for-service laboratory. In order to gain access to the core’s services an account registration form must be completed along with a service request form. Contact the Corexx or the forms or visit its offices in room 3121 on the third floor of the Harold D. West Basic Sciences Building. The PI and the FOAPAL account number to be charged must be completed. Also please provide the most accurate information on all forms. Incorrect or incomplete forms will delay the completion of your project.


TISSUE SUBMISSION

The submitting lab performs dissection, grossing, and fixation if required. Animal tissue that is being submitted for processing should have all hair removed; However, the core will perform these procedures if requested. Submit tissues in cassettes, blocks, or slides with the appropriate labeling; an example is given in the fixation section. If cassettes are submitted, please note the container solution (e.g., “Fixative, alcohol”) and preferred processing protocol (15 min., 30 min., or 1 hr per station). Fixed specimens, unless otherwise noted, are assumed to be at their end-point of fixation. Specimens for frozen sectioning are to be frozen already at -80° C. Frozen samples should be handled immediately (unless in storage at -80° C) so scheduling with the core tech MUST be reserved in advance.

All projects submitted to the core facility are completed in the order received, using the submitted date indicated on the submission form.

All submissions should be taken to room 3121 of the West Basic Science Building and handed to the Core tech. You should have an approval number accompanying the submitted specimen once your request has been received and approved.

Fixation Protocol:

  • Trim and fix the sample. Samples submitted for processing MUST be properly trimmed (not to exceed 5 mm in thickness) and fixed in the reagent of choice according to your protocol. However, the most commonly used fixative is 10% Neutral Buffered Formalin (NBF). The laboratory keeps 10% NBF, sponges, and cassettes available. Samples that have been properly fixed should be transferred into 70% ethanol alcohol and taken to the laboratory for further processing. Please call the histology staff if you need guidance on these procedures.

  • Place all submitted tissue in cassettes and properly label with a unique ID number assigned by you. Please do not fill the cassettes to capacity with tissue. This practice will result in poor tissue processing.

  • Label using the following system: JD08-####-tissue type where JD represents the first letters of the first and last name of the investigator, 00 represents the last two digits of the year, and #### is the unique ID number given by the investigator along with the tissue that is in the cassette. An example: for Jane Doe in the year 2012, 253 is the investigator’s unique ID number assigned by her and the tissue submitted is ovary. The label would be: JD12-253-Ovary

  • Use a soft #2 pencil or Secureline Marker II/Superfrost to label the cassettes. Do Not Use Sharpie Markers. The ink can’t withstand the chemicals used in processing.

  • For projects that require specially coated slides, please provide these slides at the submission of your project to the histology laboratory. Histology provides slides for routine Hematoxylin and Eosin (H&E) and IHC staining only.

  • Clearly identify all tissue containers submitted and all slides provided with the investigator’s submitter’s name.

  • Please give one week prior notification for submissions that have a large number of samples or that will necessitate frequent and continuous submissions to the histology lab.

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Frozen Section Protocol:

Samples submitted for frozen sections are required to be in clamshell molds or Cryomolds. The laboratory keeps clamshell molds, which are available by request only.

  • Place a small amount of OCT in the mold and set on dry ice to slightly cool. OCT will begin to thicken and turn hazy; add the sample and fill/cover with OCT.

  • Label the mold according to the example given for cassette labeling.

  • Place the mold on dry ice and transport it to the lab to be stored at -80°C until it is cut.

You should contact the core staff in advance for all frozen sections that are going to be performed to ensure proper handling of all samples.

Some antibodies for IHC stains require frozen sections; please contact the core tech to inquire about these particular antibodies.

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HUMAN TISSUE ACQUISITION SHARED RESOURCE
TISSUE PROCUREMENT

In order for the Core to collect and provide human research material, each investigator is required to complete a tissue procurement request form. The investigator should indicate the tissue type and amount of tissue needed, describe tissue storage and transfer conditions (e.g., media, snap freezing, and sterility requirements), and specify limiting factors in tissue collection (e.g. age, sex, race, etc.).

Application Process:

Investigators must submit a completed application. This application can be picked up at room 3121 of the West Basic Sciences Building or you can request an application be sent through e-mail. Once the application is received it will go through an approval process and you will be contacted in 2-3 business days.

If any information needed for completion of the application is missing, the application will be in pending status until we receive the necessary documents. Procurement for your project will not begin until that information is received.

Please read over the application carefully before completing and turning it in. All fees are included on the application; if you are requiring any histological services to be performed please look at the core facility fees and services for prices.