[Histonet] Re: How to troubleshoot a cytology slide with dehydration problems

[Valerie R.]

I forgot to include the complete protocol that my lab uses so you can help
me better:

1.After dehydrating in sodium chloride for 25 seconds you have to dip the
slides in alcohol 95.
2. 10 dips in water then 1.30 minutes in hematoxylin, it used to be 1 min,
then rinse in water.
3. 1 min in rinse solution
4.10 dips in water.
5. 1 min in bluing solution
6. 10 dips in water.
7. 10 dips in alcohol.
8. 10 dips in alcohol.
9. 1 min in orange solution
10. 15 dips in 95 alcohol.
11. 15 dips in 95 alcohol.
12.  3.15 mins in EA solution
13. The slides are then dipped in 3 containers of 95 alcohol (20 dips each)
14. Slides are dipped in 3 containers of 99 alcohol (15 dips in each)
15. Then they are dipped in Isoxylene which is a combination of xylene and
99 alcohol (15 dips)
16. And lastly Xylene 10 dips


When the slides do not absorb the h20 saline seems that the slides absorb
more EA than hematoxylin.  I don't know why.

On Fri, Apr 30, 2010 at 5:34 PM, Valerie R. <vrodriguez10 <@t> gmail.com> wrote:

> I am an HTL but I have to stain pap smears. In the Papanicalou procedure
> for fine needle aspirations I have to stain the slides in sodium chloride
> first for 25 seconds in total (15 seconds in one container and 10 in another
> container). This for me is the trickiest part when staining cytology because
> if slides are not well dehydrated in saline then the cells are not going to
> absorb the stains well, and the result will be a pinkish slide, when it is
> suppose to look blue-greenish.
>
> I had this problem today where I stained an entire rack of slides, and all
> the slides turned blue (which means they stained correctly) except 4 slides
> which turned out pinkish (which is a sign that they did not dehydrated
> properly in sodium chloride (h20 saline).
>
> Is there a solution to this issue. Can these slides be re-stained?
>
> I know this newsgroup is for histology only but I would like to know if
> histotech and cytotechs have encountered this issue in their labs and how
> they ended up solving the problem.
>
> Thanks in advance
>
>
>