20
adhered to the tape is rolled onto the
cold adhesive-coated slide.
FIGURE 26.
An ultraviolet flash
converts the adhesive coating into a
hard solvent-resistant plastic.
Curing the Adhesive Coating
A flash of ultraviolet light passes through the slide to polymerize the
adhesive layer on the slide into a hard, solvent-resistant plastic, tightly
anchoring the section. to the slide.
FIGURE 27.
The tape is removed.
Several options are available
Removal of Tape
The tape is peeled away leaving the still frozen section tightly bonded to
the plastic layer.
The slide can then be:
a) fixed in the
Instrumedics Aqueous Fixative
,
b)
freeze-dried or freeze-substituted in the cryostat before “anhydrous”
fixation,
c)
melted or air-dried and fixed with the fixative of your choice.
Special Fixation Protocols
Aqueous Fixative
:
THIS FIXATION SHOULD BE USED FOR ALL
HISTOLOGICAL STAINING OF CRYOJANE PREPARED SECTIONS.
This fixative melts and fixes the tissue simultaneously (direct controlled “melting”). Ice
crystal artifacts (holes) are masked. Nuclear morphology is usually excellent. Some cell
components are lost and most of the red blood cells are lysed.
1.
PREPARATION OF AQUEOUS BUFFER
Dissolve Buffer-Salt Mix in 180 ml of water.
Buffer-Salt Mix is available from Instrumedics (Cat. #AB)
Store in refrigerator. Usable for at least 6 months.
2.
PREPARATION OF AQUEOUS FIXATIVE
Add 20 ml of 25% glutaraldehyde to 30 ml of Aqueous Buffer.
Mix well.
Use at room temperature.
Discard after 1 week. (If using 50% glut. add 10 ml glut. and 10 ml water.)
CAUTION! Contains Cacodylic Acid.
Avoid contact with fingers or skin.
Do not breathe dust from vial.
3.
PROCEDURE
