Reponses to Merced and Richard Re: [Histonet] cryojane tape transfer system

[gayle callis]

You wrote:

 

Unless I'm missing something, I don't understand why people use this tape? 

It seems like a marketing gimmick to me...ol' fashion' melting of sections 

onto slides works perfectly for us...

?

Regards,

Merced

 

Merced, 

Yes,  you are missing something.  If you have ever tried to cryosection
undecalcified bone or extremely difficult tissues that simply will not
result in "ol' fashion' melting" onto a slide , then you would understand
why people use this unique cryosectioning system.  It is not some
"marketing  gimmick"  but an unique instrument  helping many laboratories
obtain frozen sections that otherwise are scrunched up,  shattered, and
destroyed.  I suggest you go to the Instrumedics website or www.alphelys.com
for a superb slide show  and learn how this instrument works before making
assumptions about a technology that serves many of us more than well.  

 

A happy, informed user of the Cryojane................

 

Gayle M. Callis

HTL/HT/MT(ASCP)

Bozeman MT 

 

As for what Richard wrote: 

 

Hi Everybody,
I was hoping to get some advice - I'm cryosectioning plant tissues and 
transferring sections to slides using the Cryojane system. However, i'm 
having problems in transferring the sections without them falling apart 
during the tape transfer. I'm fixing my tissue for 24 hours in 
ethanol:acetic acid (3:1), embedding in O.C.T, snap-freezing and then 
sectioning at between 2 and 14 microns. The sections seem to be ok but 
whenever i remove the adhesive tape from the slide a large part of the 
tissue is removed with it. As a result I lose the majority of my 
section. I've tried using both 1x and 1/2x slides (CFSA adhesive slides 
from instrumedics) but neither have given satisfactory results.
Does anyone have any suggestions as to how i could reduce the loss of 
tissue? Any advice would be much appreciated.
Thanks
Richard
 
Dear Richard, 
 
I could be the fixative you are using that causes the problem.   If the
fixative contains alcohol, the alcohol acts as antifreeze when you try to
snap freeze a tissue, animal or plant.  The alcohol may cause problems with
how the pink tape sticks to the face of the plant tissue, and allows them to
fall apart during the tape transfer.   If you rinse away the fixative, then
you should cryoprotect the fixed plant tissue with 30% sucrose before snap
freezing. vbThis will remove the alcohol.  If cryoprotection causes problems
with the final staining results, then try unfixed plant tissue, snap freeze,
Cryojane tape transfer the section and then fix the transferred plant
section in  your favorite fixative.   You may have to optimize the time in
fixative though.   
 
Other suggestions:  
 
Do a double UV flash, but wait for 15 to 20 seconds between flashes.  You
must allow the UV light source (capacitor) build up enough charge to work
properly.  This double flash seems to help polymerize the coating more
thoroughly, and the section should transfer better.  Also, the tape must be
removed at an angle across the slide,  very slowly, and inside the cryostat
(I am sure you probably do this already.) 
 
Also, try the 4X slide if you still have problems  with 1/2X and 1X slides.
You might ask Leica to send you a few to try before investing in a whole box
of these.  Contact Emmanuel Mineo, Intrumedics Product Manager
emmanuel.mineo <@t> leica-microsystems.com for the 4X slides.  Manny is a nice
gentleman who has worked with Cryojane for many years and has always been
helpful to us.   Once again, do the double UV light flash with the 4X
slides. They are gooey, but may/should hold more securely.  
 
With undecalcified bone, we use the 1/2X but do the double flash routinely
for all sections.   
 
Good luck
 
Gayle Callis