There are many cell fragments after mechanical lysis of cell suspension. How to remove these fragments? Let’s take a look at the various approaches:

1. Use dilution method. When the cells are diluted, they can also proliferate, they will become more and more numerous, and the cell debris will correspondingly become less and less.
2. There is also natural settlement. Cells settle faster than most fragments: : Move the cell suspension into the centrifuge tube, and when most of the cells sink, the upper solution can be sucked away, and then added to the culture solution to suspend the cells. This method can be used repeatedly, while each time you can observe whether it meets your requirements under the microscope.
3. Low-speed centrifuge can remove debris, generally 700g,5mins
4. When centrifuging, under the condition that the number of cells is sufficient, reduce the centrifugation time, such as 5min, 1000rpm, instead of 3min, 1000rpm, and remove the supernatant, because necrosis and debris are generally in the supernatant! This operation only needs to be performed once during the washing process before incubation!

Although there are many ways to remove cell debris, you will find that centrifugation is one of the most efficient methods！

And because plant tissues and cells have cell walls composed of cellulose, hemicellulose and pectin, it is generally necessary to grind quartz sand or glass powder with appropriate extraction solution or treat with cellulase to achieve the purpose. Bacterial cell fragmentation is more difficult, because the entire bacterial cell wall skeleton is actually a covalent bond of peptidoglycan cystic macromolecules, very tough. The common methods of breaking bacterial cell walls include ultrasonic crushing, sand grinding, high pressure extrusion or lysozyme treatment. After the tissue and cells are broken, the appropriate buffer is selected to extract the desired protein. Insoluble materials such as cell fragments are removed by centrifuge or filtration.

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