HistoGrip kicks ass over Poly-L-Lysine
I recently tried out HistoGrip from Invitrogen as a replacement for Poly-l-lysine coating for glass coverslips.
It comes as a 50X concentrate (10ml) which is enough to make 500ml of working solution.
Although it is marketed for use with attaching tissue to glass slides, I thought I would give it a try with adherent cell lines (ie HeLa) and glass coverslips.
And boy does it work better. I didn’t take pictures (too lazy), but almost immediately after seeding the cells, they began attaching to the coverslips, after a few hours they had already begun spreading, in comparison, those on Poly-L were still rounded up. There was little to no toxicity with HistoGrip, and I had a higher rate of mitotic cells remaining on the slide by the time I got to the microscope. As a test I only half coated one slide with histogrip, the coated side was confluent the un-coated about 30%. That says it all. Given the price, ease of use, and storage. I don’t think I will ever go back to Poly-l-lysine or fibronectin.
Here’s what I did:
NB: everything is to be done in sterile conditions under a culture hood.
1) Make up about 10-15ml of Histogrip in a 50ml Falcon tube, using Acetone as the dilutant.
2) Have a spare empty Falcon ready, and a 15cm tissue culture dish filled with Tissue culture grade H20.
3) Drop as many individual coverslips into the HistoGrip falcon tube as you can in about 1min. Then invert cap and invert the tube several times to ensure a good coating on each coverslip.
4) Decant the HistoGrip into the spare falcon, and then remove the coverslips from the 1st falcon and place into the 15cm plate with H20. (caution, conc. acetone will eat away at the plastic dish, so make sure you have lots of water 30+ml)
5) Keep repeating the process until you have done enough coverslips to last you a while.
6) Wash the 15cm plate 3X with H2O
7) place the coverslips on clean paper towel or blotting paper to air dry (try to separate them as much as possible to speed this up).
NB: no need to put them at 60°C, room temp is just fine !
8) Once they are dry you can store them in a dry clean falcon until you need them.
9) Invitrogen suggest you discard the diluted HistoGrip once its been used.
My guess is that you could probably get a few uses out of it. Solution goes brown after a day or so… so always use it fresh ! But in my testing I have been getting doing about 200+ coverslips each go which is enough for about 1 months of heavy experiments. That would mean that I would go thru about 120-180ml per year, and 1 10ml bottle should thus last about 3-4 years. At around 170€ a bottle, I think that re-using is probably not going to save you a whole heap of cash.