Silver staining and chromosome spreads

[kalladin]

Hi, I don't know if anyone here can help me. But I've lately been silver staining chromosome spreads for EM on plastic coated slides. However, when I do the water rinses after the staining procedure, my plastic coating just comes off and it infuriates me to no end as the whole spreading/staining procedure takes me 8 hours (6 on a good day). Does anyone have any experience and/or advice for me? I've been staining them with silver nitrate (50%) and a nylon mesh cover for 1.5 hours and adding additional stain every 15 minutes. I don't 'think this has anything to do with the temperature. I DO see some weird stuff happening that doesn't happen with normal glass slides- like how the solution bubbles up (i'm thinking some of the stain went under the plastic)?

I will appreciate any help!

Kevin

 

[Monique]

Do you buy the plastic coated slides from a company? If so, contact them and ask if they can give you advice. If you make them yourself, you need to provide us with information what kind of plastic it is and how you coat them.

 

[kalladin]

Hi Monique,

I make my own plastic coated slides. I dissolve plastic petri dishes (made by Falcon) in chloroform to make a 0.6% solution. My coating procedure involves dipping the cleaned slides into the solution for 10-15s then taking them out and immediately dipping them again for another 10s. I hang dry them in the container for a minute then remove and air dry in a dust free area. The slides turn out well and a test to see if the plastic floats off onto water turns out well.