High temperatures (and pressure) disrupt bonds within cell walls, but also denature proteins. The drawback is that it is very time-consuming. Freezingįreeze-thaw cycles work by formation of ice crystals and cell expansion upon thawing, ultimately leading to rupture. Homogenization can be performed by squeezing cells through a tube that is slightly smaller than them, thereby shearing away the outer layer (French Press) or by using a rotating blade like in a blender (Rotor-Stator Processors). Homogenizers use shearing forces on the cell similar to the bead method. This method is very popular for plant and fungal cells but comes at a disadvantage: It’s very loud and has to be performed in an extra room, otherwise you will be very unpopular.
A process called caviation occurs, in which tiny bubbles are formed and explode, producing a local shockwave and disrupting cell walls by pressure change. Ultrasonic homogenizers work by inducing vibration in a titanium probe that is immersed in the cell solution. Ultrasonictor “Amplichron®” by LKB instruments. It can be used with all kinds of cells, just add beads to an equal amount of cell suspension and vortex!
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Glass or ceramic beads are used to crack open cells – it might not sound like it, but this kind of mechanical shear is gentle enough to keep organelles intact. When the material has been disrupted, metabolites can be extracted by adding solvents. This does not have to be in suspension and is often done with plant samples frozen in liquid nitrogen. The drawback however is that the method has to be carefully adjusted as to not destroy that molecule of interest. The advantage to mechanical disruption is that no chemicals are introduced that might interfere with the substance you want to extract.
Mechanical cell disruption is really just that: forcing open the cell wall and spilling the contents.
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In Part 1, I introduced the types of cell walls out there and what they are made of – now it’s time to learn how to get through them.
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