Robust extraction workflow solutions for cells and tissues
Adaptive Focused Acoustics®(AFA®) is an advanced computer-controlled technology unique in its ability to control the amount of energy delivered to samples, such as cells. By controlling the dosage of acoustic energy delivered, it is possible to gently disrupt the cell membranes of mammalian cells (e.g., GPCR assays), or abruptly disrupt the cell walls of bacteria (e.g., total protein extraction). Cell lysis differs greatly depending on the species the cells are derived from (e.g., plant cells are significantly harder to lyse than mammalian cells). For hard and soft tissue samples, the cryoPREP® can be coupled with AFA to improve the recovery of targeted biomolecules, such as nucleic acids, proteins, and small molecules.
B. Dry tissue pulverization extraction workflow using cryoPREP and AFA
Buffer is a Critical Component
Buffer choice is a critical component for the sample preparation of primary cells and tissues. Traditionally, a buffer is selected based primarily on its lysis properties and secondarily on its biomolecule stabilization properties. Unfortunately, many protocols require harsh chemical and/or extreme physical conditions to achieve lysis, which adversely affects the quality and recovery of target biomolecules. An ideal system would initially stabilize the class of biomolecule to be extracted without a harsh and/or extreme condition. For example, a system with a rapid heat transfer from the sample to a temperature where hydrolytic enzymes are effectively inactivated, both recovery and quality of biomolecules (especially labile biomolecules such as mRNA and phosphorylated proteins) is improved. With the flash sample freezing in the thin-walled tissueTUBE followed by dry pulverization, the surface area of a sample can be dramatically increased. For example, with a 2 mm cube of tissue if converted to a 2 µm cube via the cryoPREP, the surface area increases one million fold. This increased surface area reduces the distance a stabilizing extraction buffer need to diffuse. As the Covaris lysis process is independent of buffer choice, the most appropriate buffer for the downstream analytical method is selected. The direct cell lysis and tissue processing workflows enabled by Covaris uniquely provide scientists with the flexibility to select buffers that satisfy the requirements for traditional and custom-based assays.
The controlled mechanical energy provided by AFA improves the efficiency of cellular lysis independent of buffer constituents. Therefore, AFA provides the flexibility to homogenize cells and efficiently extract proteins, RNA, DNA, Metabolites, or other targets in buffers optimized for downstream analytical methods.