How to quickly optimize homogenization protocol

How to quickly optimize a homogenization protocol for the samples you have? Many people want to get a workable protocol from homogenizer manufacturer and started optimizing from that. A few factors to consider, these are the critical factors for an efficient homogenization protocol, the sample, homogenization tube volume and temperature requirement.

Dry grinding requires much stronger homogenization tube and cap, especially with large size beads. Use the reinforced tubes for dry grinding.

Size of the beads: large size beads will generate much powerful force to break up tissue pieces quickly into smaller debris, smaller beads can then grind up the smaller pieces into a few uM particles.  Use a mixture of large and small beads will be the best of choice.

Volume of total liquid ( sample+beads+buffer) vs. the total volume of the homogenization tubes: leave at least 1/5 of the air room in the tube , otherwise the beads just slide around and be inefficient.

Temperature control: use dry ice or liquid nitrogen to keep the sample tubes chilled in the process, or use cool air to prevent sample temperature increase significantly. Remember the cold temperature will make homogenization tubes brittle and easy to break. Powerful homogenizers such as Precellys will increase the sample temperature very quickly, while Bullet Blender which uses pulse force to make beads jumping around, will not increase sample temperature that quickly.

We suggest that you run a simple protocol, and then inspect sample against light to looking for any remaining large pieces, if there still visible pieces, run a few more seconds, if not, take a small drop and inspect under microscope for particle sizes. You want the sizes are quick even, around several uM sizes.

You can also visit https://wisbiomed.com and search under tissue homogenization for more information.