The freeze, fracture, and extract integrated workflow
cryoPREP™ Extraction provides integrated solutions for processing tissue biospecimens and extracting biomarkers and other biomolecules. Tissue samples used in biomarker discovery and analytical experiments are precious, labile, and can range from very small (e.g., biopsy samples < 10 mg) to quite large (e.g., rodent organs > 2 g). Processing labile molecules and different sample masses present significant technical challenges to achieving controlled and efficient extraction. cryoPREP Extraction Systems are specifically engineered for rapidly processing tissue samples from < 10 mg up to 10 grams with continuous temperature control to ensure precious samples are properly preserved in an integrated workflow for freezing, fracturing, and extracting.
To garner relevant and useful data for biomarker discovery and other analytical experiments with clinical or pre-clinical tissue samples, it is important to properly preserve the biospecimen to capture the true in vivo biological state and native conformations. Biological processes, such as phosphatase and kinase activity, can continue long after resection of tissues or biopsy from the host drastically changing the biology of the sample; in fact, the ischemia during tissue collection can actually increase the activity of proteases, nucleases, phosphatases, and kinases. The best means to stabilize a tissue biospecimen is by flash-freezing as quickly as possible in order to cease these processes and preserve the true in vivo state. The t-PREP and tissueTUBE devices are extremely efficient at transferring heat, enabling the flash-freezing of tissue biospecimens, at least 3 times faster when compared with tissue frozen in plastic tubes.
Once samples are cryogenically frozen, it is important to maintain the low temperature and have an efficient workflow throughout the extraction process to ensure the captured in vivo state is properly maintained. In order to extract biomolecules from a frozen biospecimen it must be first broken into pieces small enough to enable efficient extraction into buffers capable of chemically stabilizing the biomarker and compatible with downstream analytical methods. The “dry-fracture” of tissue samples in the t-PREP or tissueTUBE devices with a cryoPREP™ Impactor facilitates the deconstruction of frozen tissues. The controlled impact to the sample introduces microfractures in the tissue, allowing extraction buffers to efficiently permeate the tissue mass.
The cryoPREP Extraction Systems integrate the entire tissue processing workflow, from biospecimen collection and flash-freezing through extraction of chemically stabilized biomarkers. The integrated workflow provides significant benefits including: increases in the recovery, yield, and complexity, a decrease in the hands-on time, and continuous thermal control of the sample. The cryoPREP workflow utilizes highly controlled Adaptive Focused Acoustics (AFA™) Technology, enabling release of biomarkers in buffers compatible with multiple downstream analytical methods. These integrated systems also enable optional cryogenic storage before and after the extraction process, which is extremely beneficial for rare and transient biomarkers.
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• Save time and increase sample yield with an integrated and rapid workflow.
• Preserve precious samples and labile molecules by flash freezing 3X faster.
• Maintain samples with proper cryogenic storage and continuous temperature control.
• Eliminate clean up and risk of contamination with self-contained, single-use devices.
• Experience efficient extraction with AFA Technology.
• Extract in buffers optimized for downstream analytical methods
Adaptive Focused Acoustics is a highly controllable and reproducible technology able to accurately and precisely introduce energy to tissue samples to completely disrupt and homogenize the tissue. When combined with the tissueTUBE and t-PREP advanced cryogenic tissue processing systems, AFA is extremely effective at tissue disruption and is able to enhance the extraction yield and fidelity of chemicals, biomolecules, and metabolites from tissue samples.
Chromatin Shearing Kits
truChIP™ Tissue Chromatin Shearing Kits
truChIP™ Tissue Chromatin Shearing Kit with SDS Shearing Buffer
ADME/ToxShearing for the 220 Series
Chemical extraction from tissue samples
Covaris is a technology driven company providing innovative solutions for the life sciences. If you are interested in applying these technologies to advanced tissue processing applications not listed here please contact Covaris for assistance in developing your protocols.
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J Tammam, C Ware, C Efferson, J O’Neil, S Rao, X Qu, J Gorenstein, M Angagaw, H Kim, C Kenific, K Kunii, KJ Leach, G Nikov, J Zhao, X Dai, J Hardwick, M Scott, C Winter, L Bristow, C Elbi, JF Reilly, T Look, G Draetta, LHT Van der Ploeg, NE Kohl, PR Strack, PK Majumder. Down-regulation of the Notch pathway mediated by a γ-secretase inhibitor induces anti-tumour effects in mouse models of T-cell leukaemia. Br J Pharmacol. 2009 November; 158(5): 1183-1195. Published online 2009 September 23. doi: 10.1111/j.1476-381.2009.00389 PMCID: PMC2782329
For a comprehensive list of references please see our reference section.
Figure 1: High yield and reproducible protein extraction from small tissue samples.
Total protein was extracted into 270 µl of Total Protein Extraction Buffer B from ~15 mg of mouse heart muscle using either the t-PREP™ Biomarker Extraction System or a Dounce homogenizer. Mean protein yield from 4 t-PREP samples was 182.7 µg protein/mg tissue compared to 68.2 µg protein / mg tissue with the Dounce homgenized tissue.