Cartilage staining

From:Tahir Mahmood <tahir@MIT.EDU>

Hi histonetters,

I have a question about problems i am having with staining cartilage. The
biggest issue is that the tissue has been grown on synthetic polymers, and
the polymer itself swells considerably. It is also soluble in xylene. So
"regular" safranin O staining results in 1. the overall matrix to implode,
and 2. whatever polymer was once there is dissolved (I need to see the

So to counter the effects of shrinkage etc, I fixed, froze, and infused the
tissue blocks, followed by cryosectioning. The unstained sections look
great on the slides! but then comes the staining part: the sections fall
off the +'vely charged superfrost slides during staining. 

So essentially I have 2 questions:

1. any ideas on how to keep the sections on the slides? would drying them
for a certain time at a certain temp help?
2. since these are frozen sections and i cannot use xylene, does anyone
have any ideas on how i can mount the slides without clearing with xylene?
(this is of course, once the problem of keeping the sections on is
overcome....) Could i use mounting media which is water solube (thereby
negating the use of xylene-based media), but which would not cause the
stain to bleed?

the easy thing at this point is that i am not looking to do immunostaining.
Simple Saf O would do just fine.
Any suggestions are greatly appreciated......


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