FIXATIONThe E 12 technique allows to process fixed and fresh specimens. Kaiserling solution is recommended for fixation.
FREEZING and SLICINGThe specimens are frozen in a deep freezer. For best results an ultra low deep freezer (-70 ° C) is recommended. With a special band saw body parts are sliced into 2.5 mm thick slices. After removing the saw dust, the slices are transferred to plastic grids. The grids containing the slices are placed into a steel basket for further procedures.
DEHYDRATIONThe basket of slices is placed in acetone at -25 ° C. Three or four acetone baths are used subsequently to achieve complete dehydration.
FORCED IMPREGNATIONThe basket of slices is transferred to a fresh E 12 mixture (95 pbw E 12, 5 pbw AT 30, 20 pbw, AT 10, 26 pbw E 1) and placed under vacuum for 24 - 48 hours at 0 - 10° C temperature. The vacuum is increased until 5 mm Hg is attained.
CASTINGThe basket of slices is removed from the vacuum chamber. Individual slices are placed between two glass plates. A silicone gasket is placed between the outer edges of the sheets and the clamped in position using fold-back clamps. The double glass chambers, containing the specimens, are filled with a fresh E 12 mixture (95 pbw E 12, 5 pbw AT 30, 26 pbw E 1).
HEAT CURINGFollowing 2 -3 days precuring at room temperature the glass chambers are placed in a well-ventilated oven at 50° C for another 2 - 3 days.
FINISHINGWhen curing is completed the cold glass chambers are dismantled and the sections are trimmed on a band saw. After sawing the edges are smoothed using a belt sander.