Microfluidizer® processors are the ideal equipment for cell disruption. In fact, in previous blog posts we’ve reviewed various cell lysis techniques and the advantages of Microfluidizer® technology for cell disruption. In this post, we share our tips for using a Microfluidizer® processor for lysis.
Tip #1: Do not over-mix the pre-mix.
Using a vortex mixer can entrap air in the cell suspension, which will choke the Microfluidizer® processor. This will stop the machine. The Microfluidizer® processor is not plugged but the effect is the same. Gentle agitation is all that is required to keep the cells suspended.
Ensure the cooling bath is filled with ice water during the process and refreshed as needed.
Tip #2: Match the processing pressure to the type of cell.
Bacterial cells vary markedly in their toughness. Gram negative cells like E. coli are the most commonly used and can be broken fairly easily. Gram positive cells are much tougher and should be treated like yeast or algae.
Tables 4 and 5 will help you identify the proper processing pressure based on the cell type. Run the recommended process pressure and take samples at different numbers of passes. Do not over-process. Too many passes will result in a higher degree of rupture, but at the same time protein activities can be deteriorated by too much energy input/heat generation. Over processing may also make downstream filtration and pipetting more difficult.
Chamber blockages can happen when cells are resuspended from frozen pellets if they are not all thawed, ensure complete thawing. Or when the cell concentration is too high (in that case dilute with more buffer if possible).
Tip #3: Avoid heating yeast cells to dryness
Heading yeast cells to dryness before adding them to a buffer suspension will make a tough cell wall even tougher.