Overview
A variety of tissues exist where disruption is known to be particularly difficult. Examples of these are bone, lung, or plant tissue. The Covaris AFA process works most efficiently (fastest) when a large tissue surface area is available for acoustic energy coupling. To obtain a controlled increase in surface area in a biological sample specimen of these difficult tissue types the patent pending CryoPrep™ system was developed.
Essentially, the CryoPrep is a microprocessor controlled hammer and anvil mechansim. In practice, it is an electronic solenoid developed to deliver controlled, repeatable mechanical energy (i.e., impact force) to a sample.
The TissueTube (TT1) is a highly developed tube system for use in conjunction with the CryoPrep. Simply put it is a interlocking glass / plastic film based tube that allows cryo pulverization in a very convenient and easy to use manner -- in a closed vessel. The TissueTube system has been extensively researched and exhaustively tested to ensure optimum performance every time. Indeed the polymeric film used is Kapton; most of the gold-colored material on spacecraft and satellites is Kapton. It is capable of withstanding the low temperatures of cryo pulverization and hence is the film of choice.
How the CryoPrep is Used
A sample of tissue material is placed into a tube and sealed with a plug. The tube is placed in a cryo-environment (e.g., liquid Nitrogen,dry ice or placed into a mechanical freezer) and the biological activity of the sample is quickly thermally stabilized. The sample may be stored and shipped in the tube/plug.
Prior to the freeze-fracture pulverization, a transfer tube (for example, borosilicate glass) is then attached into the top of the TT1 tube. The glass tube, TT1 tube assembly is then placed into the CryoPrep. The sample is then pulverized with a 0.5 second, one button push, the TT1, inverted and removed. The pulverized cryogenic sample is readily transferred into the glass tube, extraction buffer added and then placed immediately into either of the Covaris instruments, where acoustic disruption/extraction is carried out.
The system has gained wide acceptance for its performance and ease of use; and no cross contamination. Indeed, the TT1 tube is also used as a storage tube prior to processing. It is especially beneficial for labile biomolecules, such as RNA and enzyme substrates as the thermal control and ease of use enable fast, efficient sample processing. It is also very beneficial for hard, fibrous tissue samples as the freeze fracture effectively disrupts the extracellular matrix.
Features
- Operator safety features, such as lid and cover interlocks
- Subzero temperature and Teflon® inner lining minimize sample adherence
- Pouch material is a Kapton® laminate, an engineering grade polymer
- Total time per same <5 seconds (impact time <1 second) . Sample may be collected, stored, shipped, pulverized and transferred in the vessel to a processing vessel at subzero temperatures
- Variable impact level adjustment
- Single use, non contact eliminates clean-up time and materials
The Tissue CryoPrep system combines a single-use, consumable sample tube with an apparatus to control the pulverization impact force on the tissue sample.
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