Mechanical homogenization procedures generate lysates with characteristics different than those produced by chemical lysis. By avoiding detergents and chaotropes, many cytosolic proteins may remain intact following liberation from the cell. This is useful for protein isolation and enzyme assays. However, mechanical homogenization may simply be the tool used to rapidly disrupt cells and tissues with the use of denaturing reagents, especially during RNA isolation procedures. Regardless of the mechanical approach, whether it is to beat, grind, shear, or explode cells, they are tools that can be applied in many different ways to sample preparation.
Vortexers: Though not their intended use, vortexers are routinely used to disrupt samples. This method relies on adding grinding beads and sample to a tube and then repeatedly vortexing. Typically used for the lysis of microorganisms, vortexers can also disrupt tissues by using large grinding beads (>2 mm) made of zirconium or stainless steel.
Bead Beaters: Bead beaters moved from single tube formats to homogenizers that can handle up to 24 microfuge tubes. Microfuge tubes placed in the round rack are oscillated at high speed (up to 8000 rpm) in very short vibratory-like motion.
High Throughput Homogenizers: With the rise of high throughput screening strategies, sample homogenization became the bottleneck. To circumvent the logjam, various approaches were taken to homogenize samples en masse.
|Vortexers||Bead Beaters||High Throughput Homogenizers|
|Pros||Cost-effective||Low cost, higher throughput than vortexers and traditional lysing methods.||Highest throughput, can be paired with liquid handlers.|
|Cons||Incomplete lysis||Causes samples to heat up. Not able to lysis skin or sclera.||High inital investment. Cost of consumables|
To read more about homogenizers, see our section on "Mechical Disruption Methods" in our Homogenization Guide.
To learn more about homogenizing samples using grinding beads, see our "Bead Beating Guide: A Primer"