Hello everyone,
I wanna attach my cells on to the slide, so I used poly-lysine to coat the slide. I made poly-lysine solution in ddH2O with concentration was 1mg/ml and stored at -20oC. After that, I take this solution out and thaw, but poly-lysine could not dissolve well and there were like crystals in the solution. I tried to spin the solution and get the supernatant to coat the slide. In the beginning, the cell attach to the slide, but after several times of washing, the cells were flushed away. I also tried the washing step as gentle as possible. Do you have any idea or suggestions for this situation? And how to dissolve poly-lysine well?
Thank you so much.
Vini was right about will result in lower cell attachment sites. Commonly researcher preferred 70,000 - 150,000 of poly-lysine molecular weight for coating. But commonly, to coat the slide with poly-lysine, it has a basic guide to do it. Just flooded the cleaned slide with 1 mg/ml poly-lysine (you can choose 70,000 or higher molecular weight) in 0.1 M sodium bicarbonate buffer pH 9 for 10 sec. Drained (so the solution can be reused), rinsed with distilled water and air dried. You can find this guide in Guthrie & Fink, 2004. Guide to Yeast Genetics and Molecular and Cell Biology, Volume 1. page 128.
I hope it helps.
Hi Long, I think you can use lower molecular weight of poly-lysine (30,000 - 70,000). Lower molecular weight poly-lysine is easier to use because it is less viscous in solutions.
I would not suggest using a low molecular weight poly lysine because it will result in lower cell attachment sites. pH is the best option.
Vini was right about will result in lower cell attachment sites. Commonly researcher preferred 70,000 - 150,000 of poly-lysine molecular weight for coating. But commonly, to coat the slide with poly-lysine, it has a basic guide to do it. Just flooded the cleaned slide with 1 mg/ml poly-lysine (you can choose 70,000 or higher molecular weight) in 0.1 M sodium bicarbonate buffer pH 9 for 10 sec. Drained (so the solution can be reused), rinsed with distilled water and air dried. You can find this guide in Guthrie & Fink, 2004. Guide to Yeast Genetics and Molecular and Cell Biology, Volume 1. page 128.
I hope it helps.
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