Chambers and Coverglasses

The following contains our suggestions for the coverglasses and chambers that will optimize your fluorescence imaging and your live-cell imaging using the equipment in the CLCI:

***The main point to stress here is to always choose coverglass thickness #1.5 (0.16-0.19mm), which is optimal for most fluorescence imaging.***

Our Warner Intruments temperature controllers can accomodate both 35mm glass-bottomed dishes and 25mm diameter coverglasses.

If you would like to use the 35mm dishes with the coverglass in the bottom, we suggest you purchase them from MatTek using catalog number P35G-1.5-14-C (MatTek will send you a few

packages for free is you just want to try them). They also have dishes with larger glass inserts, pre-coated coverglasses (with collagen, for instance) and other glass-bottomed dishes like 6-wells and 96-wells.

If you want to use the stand-alone coverglasses with the gasket-sealed chamber, you need to purchase 25mm diameter #1.5 coverglasses, which can be obtained from many sources. We use Fisher Select Coverglasses no.1.5 (#NC9019858). These are a special order item and are not in the catalog. Furthermore they are quite expensive (over $500/box), but they are sorted for a consistent thickness of 170 microns.

We prepare our coverglasses by scoring them with acid, which allows many cell types to adhere more tightly to the surface of the glass. Some cells will still require the addition of a substrate (like collagen or poly-lysine) for optimal adherence, and you should determine that for yourself.

To acid-wash coverglasses:

gently shake them overnight in 1N HCl (this can be re-used several times), rinse extensively in several changes of ddH2O and autoclave them to sterilize them (this works best if you layer them on pieces of Whatman paper inside a glass petri dish - otherwise they tend to stick together).

Another option would be to use chambered coverglasses from a company like Nalge Nunc International. They make the Lab-TekII brand of chambered coverglasses with 2-8 chambers that we find useful for screening groups of cells or clones, or multiple experimental conditions.

Unfortunately we do not have an adapter for the temperature controller to use with these chambered coverglasses, so you would be limited to room-temperature and probably shorter-term examination of your cells. However, as with many resources, we could obtain an adapter for the temperature controller if enough users required it.