One of the key tasks of sample preparation is to break down the initial sample, usually a piece of tissue in the centimeter scale, to finer particles in the nano- to micro-meter scale, so target molecules, such as nucleic acids, peptides, and metabolites can be released from the tissue for further processing. Most current mechanical-based sample preparation processes cannot complete this task effectively, usually only breaking down tissue to about 100 micron particle size limit, much larger than a single molecule size level. Thus, most targeted molecules, such as DNA, RNA or protein, still trapped in the tissue mass, cannot be extracted out efficiently.

Proper tissue deconstruction is the prelude and foundation of a good sample preparation step.

The Covaris CryoPrep™ system is a patented microprocessor controlled hammer and anvil mechanism that delivers controlled, repeatable mechanical impact to a sample. This controlled impact to a frozen sample results in true freeze-fracture pulverization, efficiently shattering tissue pieces to a molecular sized level, disrupting extracellular matrix, and dramatically increasing sample surface area, which is crucial for further downstream processing. In some cases, chemical-based processes are still used with Covaris instruments. Usually, the Covaris AFA enables the chemical to reach more sample surface area and make the chemical treatment persistently efficient.

Coupled with a line of custom-designed tissue tubes, the CryoPrep offers unique advantages, including:

• The best tool for difficult tissue samples: bone, skin, corneal, muscle, lung, seeds......
• Maximum output from your "one chance to prep" irreplaceable human tissue
• True freeze-fracture pulverization (<1 second) of thermal or chemical stabilized samples
• Automated, standardized process eliminates operator variation
• No Cleaning, Non-contact, No Cross Contamination
• Ideal for labile target molecule (e.g., metabolites, proteosomes, mRNA) extraction for systems biology; the pulverized sample may be split
• Proprietary tissue tube design and pouch material for easy handling and high recovery

By using the CryoPrep, tissue disruption increased more than 15 times with RNA yield compared to conventional tissue preparatory process. . . in assays, the higher signal to noise ratio results in more visible sample complexity.

Tissue disruption using CryoPrep resulted in more than 15 times increase of RNA yield comparing to conventional tissue preparatory process...

Our customers at Genzyme were very pleased with the results, and commented: "The instrument was very quiet, gave high signal and low noise, also enabled us to see more complexity in the sample."

Small molecule drug recovery from muscle tissue resulted in higher recovery and reproducibility compared to a conventional tissue preparation process. Comparison performed by Millennium Pharmaceuticals.

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 thermally stabilized quickly. 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 within 0.5 seconds, with the push of one button, and then  the TT1 tube is 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.

Key Features

• Operator safety features, such as lid and cover interlocks
• Subzero temperature and Teflon® inner lining minimize sample adherence
• Specialized pouch material for easy handling and high recovery
• 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