Cryopreperation of tissue
A variety of tissues exist where disruption is known
to be particularly difficult. Examples of these are bone, lung or plant
tissue. AFA works most efficiently (fastest) when a large tissue surface
area is available for acoustic energy coupling. To obtain a controlled
increase in surface area for these difficult tissue types the CryoPrep
was developed.
The CryoPrep is a microprocessor controlled hammer
and anvil. In practice, it is an electronic solenoid developed to deliver
controllable mechanical energy to a sample.
The TissueTube (TT-1) 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 pulverisation in a very
convenient and easy to use manner. The TissueTube system has been extensively
researched and exhaustively tested to ensure optimum performance every
time.
How is the CryoPrep used?
A sample of tissue is placed into the tube, the tube
is sealed with a plug, which is either immersed in liquid Nitrogen or
dry ice (or placed into a mechanical freezer) and is quickly thermally
stabilised. The sample may be stored and shipped in the tube.
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 pulverised with a 0.5 second, one
button push, the TT1, inverted and removed. The pulverised 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 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.
View video of CryoPrep being used (362 KB)
download manual
Tissue range processed to date
Sciatic nerve
Brain rat, mouse, monkey - no pre treatment necessary
Colon
Liver
Rat knee
Adipose -no pre treatment necessary
Lung
Muscle
Heart
Skeletal muscle
Bone
Skin
Faeces:
Please contact Kbiosciences for more information regarding tissue mass and cryobag option.