Hi Fatumina,

what about a live fluorescent stain such as Hoechst 33342 ?

Or a label-free quantitative image processing such as PHANTAST: https://github.com/nicjac/PHANTAST-FIJI

Best regards

Article Automated Method for the Rapid and Precise Estimation of Adh...

Hi,

A while ago, I used the following protocol we published in a paper. I stained with hematoxylin. I did not recall any problem. Reference is shown.

Cell invasion assay

Cell invasion was assayed by a modified Boyden Chamber (Albini etal., 1987; Sato etal., 1994; Rochefort etal., 1998) according to the manufacturer’s instructions (Becton Dickinson). Cells were suspended in serum-free DMEM and then seeded onto uncoated filters as control inserts (8-mm pore size) or onto Matrigel-coated filters (Bioboat Matrigel invasion chambers) in 24-well plates. Fetal calf serum (5%) in DMEM was used as a chemoattractant in the lower wells. After 22 h incubation, filters were fixed in cold methanol for 15 min and stained with hematoxylin. Cells that had invaded the lower surface of the filters were counted using light microscopy at a magnification of 400. Each assay was performed in triplicate. The invasion index was calculated as follows: % invasion test cells/% invasion control cells.

Reference:

Grb2 regulation of the actin-based cytoskeleton is required for ligand-independent EGF receptor-mediated oncogenesis. Boerner JL, Danielsen AJ, Lovejoy CA, Wang Z, Juneja SC, Faupel-Badger JM, Darce JR, Maihle NJ. Oncogene. 2003 Oct 2;22(43):6679-89. PMID: 14555981

Best regards,

Subhash

You can use propidium iodide or Hoechst to stain nuclei.  This should be easily seen under a fluorescent microscope and reduce the background effects from the membrane.  Also, under higher power you can focus only on the plane of intererst.

Try Chemotaxis assay from Essen Bioscience. Currently this is the best option on the Chemotaxis assays market: http://www.essenbioscience.com/applications/cell-migration-chemotaxis/