Use
of Reichert - Jung Cryotome-2 
Hold it in position until it starts to freeze. This will attach it to the Stub & provide orientation for proper sectioning.
Add more Freezing Compound so that the entire specimen is covered. Trim away excess Freezing Compound to form a flat-topped pyramid with your specimen in its center.Dislodge the pyramid from the Stub.
Add a fresh covering of Freezing Compound to the Stub. Place the pyramid in the center of the Stub. Add more Freezing Compound around the base of the pyramid to give further support.Leave about 5 mm of the pyramid uncovered.
Place the stub in the stub holder of the Microtome Chuck.Use the Orientation Handle to rock the specimen so that its PITCH is parallel to the knife. This should produce good cross sections.
Leaf explants should be oriented parallel to the knife edge for cross sections.
This may not be a concern for stems and roots since they tend to be cylindrical
Lock the stub in place with the Orientation Handle and the Locking Screw that are part of the Microtome Chuck.
Advance the Stub towards the knife using the motor at high speed (Horizontal Fast)
Switch to Low Speed by pressing the area with two chevrons when the specimen gets close to the knife.
Do not ram the specimen into the knife! This will dislodge the specimen.
Set the thickness to 60 microns by rotating the knob next to the gauge.Make the final advance towards the knife by rotating the Microtome handle Clockwise. Each rotation will advance the specimen by 60 microns.
As you carefully continue rotating the Microtome handle, look for the first signs of sectioning. The first sections will be incomplete & curled like a scroll.
Change the thickness to 50 microns & continue sectioning until the specimen provides useful sections. The outline of the specimen will become dark when it is actually cut.
Sweep off debris with a cold brush.
The edge of the Anti-Roll Plate should coincide with the Knife-edge
Resume sectioning. Sections should collect underneath the anti-roll plate.
Collect no more than 3 sections! Place a cold microscope slide next to the knife holder, on the left side of the Anti-roll Plate.Specimens may adhere to it or they may stay behind, or they may do both. Do NOT Panic!!!!!
If they adhere to the anti-roll plate, place the microscope slide underneath the plate and use a cold brush to gently dislodge them so that they fall on the slide.
If they stay behind, use a cold brush to gently sweep them onto a microscope slide.
Carefully lift the slide out of the chamber. The sections will fall off the slide if you move it too swiftly!
The sections will start to melt. At this point you can do one of the following
a] add liquid Freezing Compound which will act as a mounting medium, the add a coverslip.
b] add Water to dilute the Freezing Compound so that the sections can be Stained. The freezing compound interferes with staining.
c] let the sections melt very slowly for delicate specimens, add a coverslip and photograph right away.
Cryosections are very delicate. They are especially damaged by sheer forces. These occur when you rotate the coverslip AFTER it has settled. Consequently, add the coverslip carefully and correctly!
Specimens may fall off the block during sectioning.
a] Simply put some fresh freezing compound on the stub and press your specimen onto it.
b] Be sure to withdraw the stub from the knife & repeat the alignment of the specimen to it.
