AFA™ is unique in its ability to control the amount of power applied to cells. This means it is possible to gently disrupt a cell membrane (e.g., in a G-protein coupled receptor assay) or abruptly disrupt the cell (e.g., a total protein prep). Indeed, it has been shown to achieve sub-cellular fractionation by using differential "mechanical lysis" conditions.

Cell lysis differs greatly depending on which cell species is being disrupted (e.g., a plant cell is significantly harder to lyse than a mammalian cell). Consequently, the buffer is a critical component when using the AFA processes. Typically, researchers choose a buffer by acknowledging the stabilization characteristics for their target molecule, rather than for the lytic capabilities. The AFA process provides the mechanical energy to also improve the efficiency the buffer constituents. For example, lower levels of surfactants may be used reproducibly to isolate cellular components with the AFA process.

Covaris has shown successful lysis of the list of organisms below. The researcher should bear in mind the requirements for this application, and consult with Covaris should they have a specific cell lysis application in mind.

To date the following organisms have been successfully disrupted:

• E. coli
• A variety of tissue culture derived cells (e.g., HeLa)
• Fungal spores
• Yeast
• Plant Cells

Unique Features and Benefits

Feature	Benefit
Non-contact	Sample is not contaminated
Controllable mechanical energy	Dissolution is controllable for all application areas Increased biological activity in preparations
Electronic driven	Highly repeatable
Automated – E series	Walk-away sample disruption
Isothermal	No heat generated Lysis can be controlled at temps of 4°C to ­40°C
Fast	Many samples can be processed in very short time frames
Samples are closed tube	Safe - no aerosol risk No sample cross-talk No clean up, or decontamination
Buffer independent	Some processes may be carried out in the absence of "traditional lysis buffers"
Scalable	Volumes from 10µl - 10mls
Automation enabled	E-series instruments can process microplates or racks of tubes

Preparation Types

A number of different cell lysis preparations have been conducted. For more information please contact Covaris. One example is the lysis of Saccharomyces cerevisiae (yeast). The data presented below shows complete yeast cell carcasses after AFA treatment and a mixture of intact cells and carcass fragments after conventional glass bead treatment. The application was for intracellular protein extraction.

Yeast cells intact	Complete cell carcasses after AFA treatment
Mixture of complete cells and carcass fragments after conventional glass bead treatment.