I am looking for a protocol (cheap and effective way) that I could use to isolate multiple types of cells and be able to sort them out. For instance, I want to isolate multiple sub populations of interstitial cells in the aortic valve.
Any help/recommendation would be greatly appreciated. Thanks.
Hi,
After the mechanical/enzymatic digestion of your tissue, you can do Antibody staining based on the surface markers of your cell types. Then you proceed with FACS sorting. This is the best way to go for separating cell of different types/subtypes. Anyway, there are also way to enrich for some populations with MACS approach.
Best,
Michail
Dear Sana,
Based on IHC or other stains such as H&E, you could identify your cells of interest under the microscope and use laser microdissection (e.g. MMI CellCut) to isolate the cell populations directly from tissue sections. The software tools allow you to define different groups of cells which can then be collected into different tubes for subsequent analysis. I like this technology as you maintain the spatial information.
Alternatively, you could digest the tissue to create a cell suspension, stain your cells based on the markers of interest and use a micromanipulator (e.g. CellEctor) to pick your cells. Again, you could define group of cells and sort the them automatically into different tubes.
FACS and MACS are very well suited for a large number of cells. However, if you are focussing on the selective isolation of a smaller number of cells (up to a few thousands) I would recommend the CellCut or CellEctor system. Both systems can go on one platform, so you could work with tissue and cell suspension.
I hope this helps!
Kind regards,
Heidi
Just seeing this thread now, but I am going to answer this here in case it is helpful to anyone who might stumble upon this .... You could look up this website: www.rnassist.com. This product called vivoPHIX enables tissue fixation and then dissociation for tissues, prior to downstream applications !
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Materials
- Inorganic Materials
- Metals
- Alloys
- Steel