Carnoy's fixed tissue - Photomicrographs and Method

From:ncragg

Thank you for all your replies  - I am very grateful  and hopefully I will 
be able resolve this problem.  I have attempted to post a couple of 
photomicrographs on the website to demonstrate the nuclei; 
tonguecarnoys.jpg & gutcarnoys.jpg   It may not be as good quality as you 
are used to as I have only been practicing histology for a few weeks!  The 
photomicrographs don't look exactly like what you see down the microscope, 
but this is also the first time I've used the Spot Advance Camera!!

The protocol used for the tissue shown in the photomicrographs was as 
follows:-

Fix in carnoy's for approx 1 hour, then transfer to 70%  before processing. 

1	70	30 minutes
2	90	30 minutes
3	95	30 minutes
4	100	1 hour
5	100	1 hour
6	100	2 hours
7	100	2 hours
8	Xylene	1 hour
9	Xylene	1 hour
10	Xylene	2 hours
11	Wax	2 hours 30 minutes
12	Wax	4 hours

Initially I used shorter programs, but on finding these black/brown nuclei, 
we decided to increase the length of time in 100% and Xylene to see if that 
would resolve it, which is how we arrived at the protocol above.

I am going to try rinsing in 95% or 100% before processing as Gayle 
suggested.

FAO: Terry Marshall - Nice to hear from someone over this side of the 
pond!!  I will post you a slide if that helps!?


Kind regards,

Nicola Cragg
Epistem Ltd.
Manchester, UK




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