: [Histonet] Xylene substitute for processing GI biopsies

[Teri Johnson]

Hi Lori,

Pretty much any xylene sub will work for processing tissues and staining. You may need to add additional time in the "xylene" steps, since the action of the subs isn't as quick as xylene.
In addition, the substitutes are more intolerant of water. How humid is your lab? If it's dry, then you may not have to make any additional adjustment. Otherwise you may need to make sure your final absolute alcohol is fairly anhydrous or water will collect in the bottom of your container, potentially causing problems with processing.

Molecular sieves can be used to keep that from happening, or you can add additional containers of absolute alcohol (and decrease the time accordingly in each one) so there isn't as much water carryover by the time it reaches the first container of substitute.

Some people are sensitive to the orange smell of the Limonene subs. I always liked it, but it gave headaches to my supervisor when we were trying it. Plus it is more oily than xylene so your slides might take longer to dry after coverslipping.

If you coverslip out of xylene sub, you need to you have a mounting medium compatible with the substitute. Check with the manufacturer to see what they recommend.

Teri Johnson
Manager, Histology
Genomics Institute for
Novartis Research
Foundation
San Diego, CA
858-332-4752