I need to perform cell migration assay using transwell inserts of 8 micron pore size membrane and matrigel. What is most preffered membrane for such experimets: Polycarbonate, PTEF or PET?
Hi,
It depends on the type of experiment. PET provide better cell visibility under phase contrast microscopy and more fluorescence imaging quality; and allows assessment of cell viability and monolayer formation. PTEF are not tissue culture treated so they have low binding properties and they are not the best for fluorescence imaging. PC membranes are better for barrier assays but not for imaging. In most papers I have seen they use PET membranes and it works for me.
Here, there are two useful links:
https://www.warneronline.com/Documents/uploader/Corning-Snapwell-Transwell%20Instruction%20Manual1.pdf
http://csmedia2.corning.com/LifeSciences/Media/pdf/transwell_guide.pdf
Hi Pooja
I have been using polycarbonate membrane and it has been working well (Corning Star 8uM).For staining, crystal violet works fine.
Hi Pooja,
I was using Transwell inserts (polyethylene terepthalate (PET) track-etched membrane-diameter 6,4 mm; 8 µm from Falcon (BD Labware, USA). They worked perfectly for me.
Hi, I would like to add one thing here, based on the type of cell, staining method and gene of interest/phenomenon I should use choose the type of transwell. I assumed here this is just migration not invasion (no need for coating with fibronection and endothelial cells) assay. PET is working well regardless the vendors.
Best,
Zack
Dear Pooja,
I would suggest you go through the following link to understand what exactly you want to use.
https://www.bdbiosciences.com/documents/webinar_2008_12_fluroblok.pdf
I use Corning® BioCoat™ Fibronectin Inserts with 3.0µm PET Membrane, 12 inserts/plate for endothelial cell migration assay. It works fine for me. Hope you have good filter based fluorescence reader with bottom reading option like Envision.
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