If you are conducting experiments with a specific cell type, the first procedural step will usually involve preparing the cells from a tissue of interest. This entails lysing the cells to release intracellular contents, usually nucleic acids and/or proteins. The process may be accomplished in a number of ways, each conferring varying degrees of effectiveness, cost, and time. High pressure homogenization is known to be one of the more effective methods of cell lysis. Below is a list of factors to consider as you create a tissue homogenization protocol, intended to enhance the quality and amount of lysate.
- Protein location
Cellular location plays a huge role in determining the best way to approach cell lysis. For example, certain proteins are located in specific organelles. Lysis methods that release the contents from every compartment make it incredibly difficult to achieve high yield of a protein of interest. In contrast, optimization of methods that are able to release contents from specific structures makes the process easier, and enhances product isolation.
- Size of samples to be processed
Every homogenizer is suited to a specific range of sample sizes. For example, bead mills tend to homogenize the smallest sample sizes (2 mL or less), while industrial-scale homogenizers can process many liters at once. Being able to predict the range of sample sizes you will be mostly working with before purchasing a homogenizer is ideal. However, if you already have a homogenizer but your sample is the incorrect size, you still have options. Samples that are too large can be cut into smaller pieces, and samples that are too small can be diluted to a volume that is better suited to the machine.
- Tissue type being homogenized
The type of tissue your cells come from will largely dictate the sensitivity of your protocol. Soft tissue, such as adipose, muscle, and connective, are not difficult to disrupt. They therefore require a sensitive process that lyses cells without destroying the proteins of interest. In contrast, tough and fibrous tissue will need to be paired with a powerful process that is able to break through rigid cell structures.
Pion: The Homogenizer Advantage
Use of high quality cell lysis equipment will provide numerous benefits for applications like tissue homogenization. And there are plenty of companies on the market to select your equipment from. However, the lysate can be of higher quality and more even consistency when run through top-shelf equipment, most frequently in the form of a high pressure homogenizer. Pion's BEE brand technology is trusted by researchers and lab managers around the world. We deliver an array of key benefits, such as production of nano/micro emulsions and dispersions and lipids and suspensions; these can be used for applications such as injectables, targeted drug delivery, inhalants, time release, anesthetics, and importantly, vaccinations.
In addition, we have extensive experience in the challenges that our customers face as they transition from concept, through to R&D, clinical trials, all-important FDA approval and finally, to manufacturing.
Learn more by visiting our products.