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 (for example in a G-protein coupled receptor assay) or abruptly disrupt the cell (for example a total protein prep). Indeed, it has been shown to achieve sub-cellular fractionation by using differential "mechanical lysis" (AFA) conditions.

Cell lysis differs greatly depending on the cell species 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 for its stabilization characteristics for their target molecule, rather than for its' lytic capabilities. The AFA process provides the mechanical energy to also improve the efficiency of buffer constituents. For example, lower levels of surfactants may be used, reproducibly to isolate cellular components with the AFA process.

Covaris to date has shown successful lysis of a number of organisms as listed below. The researcher should bear in mind the requirement of the application and consult with Covaris should they have a specific cell lysis application.

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, nor 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 conduct Covaris. One example of this is in the lysis of Saccharomyces cerevisiae (yeast). The data presented below shows yeast cells intact, complete cell carcasses after AFA treatment and a mixture of complete cells and carcasses 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.