Cells are P. falciparum freed from RBC's. This should be a quite fast method since I want to study transport processes.
Thank you.
Hi Praveen,
I want to incubate saponine lysed P. falciparum infected RBCs with different radiolabled substrates and check after certain time points the uptaken amount of radiolable.
Hello Janis,
I am not sure what is the downstream treatment of the isolated parasites. Centrifuge at lower g. you can try standardizing the lowest speed for saponin treated cultures without uptake experiment, and then perform your experiment.
and in case you just want to replace culture medium with another medium then: Pass the isolated parasites on magnetic column in presence of magnetic field and elute in the absence of magnetic field in desired medium.
I am not sure if there is some report about some density gradient method for this purpose.
Praveen
Hi,
what is excatly your cell separation problem?
If you want to sort RBC from falciparum, you can used density gradiant for example.
I also develop some SdFFF (sedimentation field flow fractionation) prototypes which are used as cell sorter, without labeling.
The separation is based on the intrinsic biophysical parameters such as size, density, shape and deformability
Hey thanks for the answers! My problem is that I have to stop the radiolable substrate uptake. This takes place in an aqueuos solution. Normally I wash and filter my cells (usually yeasts) through GFC filters. I'm afraid that the parasite cells will not stand this treatment and break. This would result in an unpredictable loss of radiolable. The normal procedure for substrate uptake measurements in P. falciparum is a centrifugation step through an oil phase which separates the cells from the aqueous phase. But I have some problems with this method and wanted to try something else.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Molecular Biological Techniques