Cell disruption

Cell disruption is a method or process for releasing biological molecules from inside a cell.

The production of biologically interesting molecules using cloning and culturing methods allows the study and manufacture of relevant molecules. Except for excreted molecules, cells producing molecules of interest must be disrupted. This page discusses various methods.

A common laboratory-scale mechanical method for cell disruption uses tiny glass, ceramic or steel beads mixed with a sample suspended in aqueous media. First developed by Tim Hopkins in the late 1970s, the sample and bead mix is subjected to high level agitation by stirring or shaking. Beads collide with the cellular sample, cracking open the cell to release intercellular components. Unlike some other methods, mechanical shear is moderate during homogenization resulting in excellent membrane or subcellular preparations. The method, often called "beadbeating", works well for all types of cellular material - from spores to animal and plant tissues. It is the most widely used method of yeast lysis, and can yield breakage of over 50%. It has the advantage over other mechanical cell disruption methods of being able to disrupt very small sample sizes, process many samples at a time with no cross-contamination concerns, and does not release potentially harmful aerosols in the process.

In the simplest example of the method, an equal volume of beads are added to a cell or tissue suspension in a test tube and the sample is vigorously mixed on a common laboratory vortex mixer. While processing times are slow, taking 3-10 times longer than that in specialty shaking machines, it works well for easily disrupted cells and is inexpensive. process times can be improved by using a bead dispenser to load the beads into the vials or plates. The company LabTIE produces Bead dispensers that dispenses the beads in a grid in only 8 seconds.

In most laboratories, beadbeating is done in sealed, plastic vials, centrifuge tubes, or deep-well microtiter plates. The sample and tiny beads are agitated at about 2000 oscillations per minute in specially designed vial shakers driven by high power electric motors. Cell disruption is complete in 1–3 minutes of shaking. Machines are available that can process hundreds of samples simultaneously inside deep well microplates.

Successful beadbeating is dependent not only design features of the shaking machine (which take into consideration shaking oscillations per minute, shaking throw or distance, shaking orientation and vial orientation), but also the selection of correct bead size (0.1–6 mm diameter), bead composition (glass, ceramic, steel) and bead load in the vial.

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