[Histonet] Superfrost Gold versus Superfrost Plus Charge

[gayle callis]

You did not say whether the tissues were prefixed with formalin or fresh
tissues, frozen then cryosectioned?   If prefixed, cryoprotected, frozen and
cryosectioned, the Plus Gold should work.  They are certainly more
expensive!   A colleague uses Plus Gold  for prefixed, cryoprotected frozen
sections destined for immunostaining.  She sections, then air dries for a
time.  You may need to determine the time of drying, even if the tissue is
fresh.  There are package inserts that tell you HOW to use these slides,
both Gold or Plus Charge, at least the ones manufactured by Erie have an
insert.  The Gold slides should be dried flat, but that may be due to
contact with a room temperature surface e.g. tray.  To help you air dry,
purchase a cheap, small fan and dry any FS on Gold or Plus Charge - make
sure  the air blows pretty much directly onto the sections whether you lay
them flat or slanted to be in airflow.  You will hasten the drying time and
dry more efficiently even in a 30 minute time frame.   Try and get a Gold
Plus samples from Erie/Thermo Scientific rep in your area, hopefully they
still do this. 

 

Superfrost Plus Charge (Erie trademarked)  works very well with fresh tissue
frozen sections, followed by fixation. If we section fresh tissue, air dry a
minimum of 30 minutes, solvent fix and stain, we do not lose sections but we
also do NOT use an automated staining system.   However, if you fix with NBF
and use a retrieval method (MW or enzyme digestion) the prefixed or NBF
fixed fresh frozen sections may lift off.   

 

We  never use any kind of gelatin or glue as an adhesive for immunostaining
work, just the Plus Charge (trademarked, meaning Erie manufacturered).  I
have heard the Mercedes coated slides work very well with prefixed tissue
frozen sections.  You may want to try those too.   

 

Since frozen sections are notorious for being more delicate, they cannot
withstand some mechanical manipulations such as the reagent dispensing on a
stainer - maybe too forceful???  Maybe that can be adjusted so it still
allows good flow but not quite so harsh??  

 

If you cut fresh frozen sections, air dry, fix with 4C acetone for 10 min,
dry to evaporate solvent and stain, you can stain.  A common complaint is
poor morphology, but you can post fix a totally IHC frozen section in NBF
for 10 min, rinse, then counterstain with hematoxylin.  Dr. Chris van der
Loos does this with great success and improves the morphology on his section
by doing so.  

 

Good luck

 

Gayle M. Callis

HTL(ASCP)HT,MT

Bozeman MT 59715