[Histonet] Spectra

Gudrun Lang gu.lang at gmx.at
Tue Apr 21 03:15:06 CDT 2020


Hi!
Do you use your own reagens or the Leica Spectra dyes? We have house-made
ones.
Too faint eosin staining can be rendered more intense with a few drops of
acetic acid into the eosin to reach a pH about 5. Second, it is often a
prolonged washing after the eosin that causes excess stain to be lost.

Our protocol is 1 min in 2% aequoues eosin (pH 5,5), rinsing in running
tapwater for 1 min, then 1 min in 96% ethanol,  2x 100%, xylen. The times
for rinsing in water and 96% have to be "exact" in the protocol. In 100% and
ethanol a prolonged stay doesn't matter.
We optimized the turnaraound time with two identical protocols for HE, that
use two staining dishes each for hematoxylin and eosin. The rest of the
dishes is used by both protocols. Except of the former mentioned stations,
the rest is "not exact". So the Spectra has more "ways" to find an optimal
protocol.
The all-over staining time is also about 60 min, due to 25 min in the oven
and the progressive hematoxylin (10 min). Usually we feed the stainer
continuosly  in 15-20 min steps, as the racks are filled after cutting. And
the racks are ready in the same pattern.

Here, we have no problems with too fast lowering of the reagens-level.

Hope, this helps.
Gudrun

-----Ursprüngliche Nachricht-----
Von: warda hassan via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Gesendet: Sonntag, 19. April 2020 11:37
An: histonet at lists.utsouthwestern.edu
Betreff: [Histonet] Spectra

Good Morning to all

Thanks in advance to all who would help with their feedback on the
automation of H/E staining,
Initially we have installed Spectra-Leica for H/E
The feedback from pathologist is the stain is bit on bluish and eosin is
weak
We did altered and reached to 2 min of Hematoxylin and 11 min Eosin
But unfortunately the stain consistency is not stable as after every 2-3
racks level of reagents reduces so we top up as we do the intensity of it
differs
Second issue is Turn around time, each rack takes 45mins, so tried adding
up new protocol as copy but the machine keeps a gap of 12mins in picking up
the slides for staining !
If anyone of you have experienced with Leica automation
Please your feedback on above would be of a great help
Thank you so much
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