Protocols

Fixed Tissue

1. Tissue should ordinarily be no greater than 3 mm thick for proper fixation. Larger tissues maybe fixed for one hour, then sliced part way through like a loaf of bread to promote penetration of the fixative.
2. The usual fixative for animal tissues and paraffin processing is either 10% Neutral Buffered Formulin (NBF) or Paraformaehyde(PF). Alternative fixations for immuno-staining are usually based upon coagulative fixatives like ethanol, methanol or acetone or aldehyde with permeabilization and antigen retrieval. We can work with investigators to determine the optimal fixative (s) for their projects or if staining problems are encountered.
3. Place tissue in standard histology embedding cassettes (Fisher # 15-182-500E) labeled with a special histology solvent proof pen (Fisher # 1450-20-FSC).
4. The cassettes should be identified using the following system: AB00-1234. Where AB represents the first letter of the first and last name of the investigator, 00 represents the last two digits of the year, and 1234 represents a sequential log number that the investigator maintains to track his samples. An example for John Smith's sample number one of the year 2002 would be JS02-0001
5. Tissue should be fixed overnight dependant upon size and tissue type, then placed in 70% ETOH for storage and shipment. Do not fix for more than 48 hours.
6. Place the samples with fixative or alcohol into a Kapak/Scotchpak heavy-duty heat sealable pouches. Press flat to remove air bubbles and seal with hot iron about 2/3 up the bag. Double seal by sealing at the very top of the pouch.
7. Samples in alcohol maybe shipped via US mail, samples in formaldehyde must be shipped by a carrier approved for hazardous materials to the following address:

   
   Van Andel Research Institute
   Analytical, Cellular, and Molecular Microscopy Lab
   333 Bostwick, NE
   Grand Rapids, MI 49503
   Phone: 616-234-5265

Frozen Tissue

1. Fresh tissue should be frozen in Tissue-Tek II Cryomolds (Allegiance #M7144-11) with Tissue Tek OCT compound (Allegiance #M7148-4) on dry ice.
2. Label the cryomolds with a laboratory marker using the following system: AB00-1234. Where AB represents the first initial of the first and last name of the investigator, 00 represents the last two digits of the year, and 1234 represents a sequential log number that the investigator maintains to track his samples. An example for John Smith's sample number one of the year 2002 would be JS02-0001.
3. Place the cryomolds in a Kapak/Scotch sealable pouch; include the investigator name and tissue information. Seal the pouch and store on dry ice or in a -80 degree C freezer until shipment.
4. Ship the frozen samples on dry ice using an overnight carrier to the following address:

   
   Van Andel Research Institute
   Analytical, Cellular, and Molecular Microscopy Lab
   333 Bostwick, NE
   Grand Rapids, MI 49503
   Phone: 616-234-5265