Essential Methods of Plant Sample Preparation for Light Microscopy - Plant Cell Morphogenesis: Methods and Protocols - page 18

Fig. 3 Detail of cryotome head with chuck and object. In the lower part of the
picture is the anti-roll plate with section underneath ( a ). To pick up the section,
rise up the anti-roll plate and place the slide very close to the section (resting the
corner of slide on the blade holder helps). The section “jumps” on the slide and
melts on its surface ( b - d ). Do not press the slide on section, as it would freeze
onto the blade holder. ( e ) Slide with two collected sections

8. Fix the chuck into holder and adjust its axial position with
microtome head according to the blade. Approach slowly with
objects towards the blade. Do not trim too quickly as damage
may be caused to blade and object can break off.
9. Set the position of anti-roll plate parallel with blade edge. The
edge of the anti-roll plate should be close enough to the edge
to allow emerging sections to slide underneath (up to 0.5 mm)
but not too close to crush the block. It is not easy to adjust
position visually, so while very gently touching the top of the
plate, one should feel the cutting edge. Be careful not to hurt
your fi ngers! Cut few sections (it is reasonable to start with
section of 15

m) and further correct the plate position. If the
section rolls in front of the plate, move it slightly up. If the
plate is too high, object touches the plate lightly in passing
(can be sensed with fi nger touching lightly the plate holder)—
lower the plate.
10. Cut several sections to make smooth block surface.
11. Cut the sections; they run individually or in row of few under
the plate (Fig. 3a ).
12. Pick up sections with brush chilled in cryotome chamber or
collect them with subbed (adhesive coated) and marked slide
of room temperature. The slide should be approached very
closely to sections but not direct touching them. Sections will

μ