I want to prepare cells for fluorescence microscopy staining. My cells are adherent. Do you think I can do it in chamber slides or do I necessarily have to do it in coverslips?
Hi Esmeralda,
Chamber slides and coverslips both are good for fluorescence microscopy. But use of coverslips is much cheaper than the chamber slides.
Good luck!
Arun
Thank you Bart, it is indeed a good suggestion. I'll see if I can have them in Mexico. How many cells can you seed in those chambers? Mine are big (macrophages).
I've used chamber slides and coverslips for IF of cells. We usually coat the chamber slides with fibronectin before using them.
Hi Esmeralda
Whatever you are going to use cover slip or chamber slide, it is very important to get sharp and good pictures after performing your IF staining. So, if you are going to use coverslip, you need to adhere (mount) it to a slide and if you are going to use chamber slide, also you need to cover the cells by a cover slip and in both cases a special mounting medium is needed. So, the thickness of the cover slip in both cases is very important to decrease the aberration and increase the image resolution and intensity. Finally, i advice you to try cover slip # 1 for fluorescence microscope and cover slip # 1.5 for routine staining such as Crystal violet, Giemsa, or Hematoxylin and Eosin. .
Good luck
Hello,
Coverslipping the slides is especially important when doing IF due to photobleaching. So, most likely you will need to coverslip them and make sure to use a mounting medium that will minimize your photobleaching effects. DAKO makes a nice aqueous based mounting medium that even has DAPI (nuclear marker) already in the mix, so you can eliminate this step if you'd like to see your nuclei as well.
Best,
Toby
As already mentioned, you can use both, chamberslides as well as coverslips for performing IF on adherent cells.
Chamberslides are of course more pricy than coverslips, however, they have several advantages: From the handling procedure it is much easier to do your IF on chamberslides than on coverslips (the slips are very fragile what means that frequently some of them get broken and, as you have to work with forceps, on every handling step you destroy some of the adhering cells).
Another big advantage of using chamberslides with four or eight chambers is that you need much less cells for your assays compered to coverslips.
It’s also an advantage that you can stain different chambers with different antibodies what means that the samples for your controls and the samples for the proteins you want to investigate were grown under exactly the same conditions.
And, finally, there are chamberslides available, which are not made from glass but from a special plastic. These plastic slides are especially useful when working with adherent cells growing very well on plastic, like 12-well plates, but making problems when cultivated on glass.
Chambers are usually for easing live studies on inverted microscopes. Even if they are described as embeddable, i had a negative experience with some. Might be others works. ( i am speaking about chambers on microscopic quality glass coverslips.) coverslips in an appropiate container are more meaningful and simple. Use glass coverslips with diameter 12 mm; they fit into a 24-well plate. Of course, this is a handyman work. Starting by plating cels in drops usually. Anyway, let's go ahead. :) end-solving is a 24-well plates with removable glass coverslip as backs.
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