There are many methods for mounting slides. What is important isn't the exact method at left, but what it provides the cells between the coverslip and slide: space. The four dabs of petroleum jelly or dried nail polish keep the coverslip and slide far enough apart to prevent the sample in between from getting flattened. Also, an anti-fading agent in the mounting media is essential.  To prevent sample movements, the edges of the coverslip must be fixed in place with nail polish, super-glue, velap, etc. if an oil immersion objective is used for imaging.

Other methods include using number 0 or 1 coverslips cut in half and placed at the edge of the coverslip with the sample to provide a bridge. Some people prefer a beeswax/petroleum jelly mixture to nailpolish. Some people prefer Vectashield or Dabco to the n-propylgallate.

Our confocal microscope is on an inverted Nikon. Keep in mind that the slide and coverslip must be turned upsidedown. Please only put one coverslip per slide.

Please see our page on dishes and chambers for more information.

Please see our suggestions for immunofluorescence staining for more details on preparation.

Sender: Confocal Microscopy List     From: Rosemary White
Subject:  slide sealants and cell viability, GFP, etc.
To: CONFOCAL@LISTSERV.ACSU.BUFFALO.EDU

Dear all,

I would just like to emphasize the usefulness of Valap,
which I've been using  to seal slides since student days.  On
occasion, I've wanted a harder sealant or one that adhered better to wet
slides, and there are a number of dental waxes of varying hardness that
work very well for sealing in living tissue - like the dental wax used to
seal knife boats when sectioning.  One we've used recently is Kemdent
sticky wax, made in the UK by Associated Dental Products - sets to hard
candle wax consistency.  We've also used small drops of this dotted on the
corners of a coverslip to prevent squashing of fragile specimens, like
protoplasts.  A soft sealant, consistency of plasticine, that also works
well is Surgident periphery wax from Heraeus Kulzer, Dental products
division, made in USA.  Marvellous stuff!

Rosemary White   Microscopy Centre   CSIRO Plant Industry
GPO Box 1600   Canberra, ACT 2601   email: r.white@pi.csiro.au