Cell Lysis
AFA is unique in its ability to control the amount of power and ultimately
the dissolution state of cells. This means it is possible to very gently
disrupt a membrane (for example in a G-protein coupled receptor assay)
or greatly disrupt the cell (for example a protein prep), indeed it is
theoretically possible to achieve sub-cellular fractionation by the use
of differential lysis conditions.
Cell lysis differs greatly depending on the cell species being disrupted.
In other word a plant cell is significantly harder to lyse than a mammalian
cell.
To date the following organisms have been successfully disrupted.
E.Coli
A variety of tissue culture derived cells (eg. HeLa)
Fungal spores
Yeast
Plant Cells
E.coli cell lysis using 9 recombinant proteins
Lysis comparison
Bands indicate comparable solubility rate with conventional methods
Bands indicate chemical lysis solubilise more insoluble membrane proteins than Covaris AFA
Yeast cells lysis
A number of different cell lysis preparations have been conducted. One example of this
is in the lysis of Saccharomyces cerevisiae (yeast). The data presented
below shows yeast cells intact, complete cell carcases after AFA treatment,
and a mixture of complete cells and carcases after conventional glass bead
treatment.
Yeast cells intact
Mixture of complete cells and carcass fragments after
conventional glass bead treatment.
Complete cell carcasses after AFA treatment
Frozen
cell pellet being lysed in real time (206 KB)
Features & 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 |
| Isothermal | No heat generated. Lysis can be controlled at temps of 4°C – 40°C |
| Fast | Many samples can be processed in very short time frames |
| Samples are closed tube | Safe – no aerosol risk |
| Buffer independent | Some processes may be carried out in the absence of "traditional lysis buffers" |
| Scalable | Volumes from 200µl - 10mls |
| Automation enabled | E-200 can process microplates or racks of tubes |
Presentations
Final report Industrial Placement student
Janice Cheung
Evaluation of the Utility of Adaptive Focused Acoustics Within a Discovery Research Department
Global Protein summit
London 2006
An Investigation into the Utility of non contact isothermal Adaptive Focused Acoustics in Bacterial Cell Lysis
