Frozen section
Cutting the frozen section

Author: Jessica Wallace (see Authors page)

Revised: 26 July 2017, last major update October 2011

Copyright: (c) 2002-2017, PathologyOutlines.com, Inc.

PubMed Search: Cutting the frozen section

Cite this page: Cutting the frozen section. PathologyOutlines.com website. http://pathologyoutlines.com/topic/frozensectioncutting.html. Accessed September 24th, 2017.
Procedure
  1. Once the chuck is in position, there should be a manual or an automatic advance option to move the block close to the cutting blade

    Tissue embedded within OCT

  2. Fully face the tissue by using a trim setting on your cryostat; if you do not have this setting, then an advance button should be available, which should be pressed each time before one full revolution of the instrument's wheel
  3. If wells are used to freeze the blocks, then the tissue should be on an even plane and the tissue will be faced faster
  4. To polish the tissue, avoid advancing the cryostat or deselect the trim setting on the cryostat and turn 10 - 15 times
  5. As you cut the tissue, anchor the tissue to prevent folding or curling; this can be done with an anti roll bar (a plastic plate attached to cryostat) or by using a precooled paintbrush with stiff bristles and a wide gripping surface
  6. The brush should be held like a pen with your left hand at an angle
  7. You can rest your fifth finger on the stage for stabilization
  8. Cutting the brushes' bristles at an angle can aid in the brush meeting the tissue flat over its length because you will hold it at an angle

    Left: brush with angled tip; right: holding the brush

  9. Turn the wheel with your right hand in a continuous motion without stopping; avoid speeding up or slowing down
  10. Avoid stopping the wheel at the beginning of the section, slowly grabbing the tissue and then resuming wheel revolutions; this can cause artifacts such as variation in section thickness and tissue folding
  11. Move the brush as the chuck moves towards the blade; your brush should move down in pace with the chuck

    Riding the block: as the block descends
    toward the brush, the brush keeps pace
    with the block by gently resting on the
    bottom 2 - 3 mm of the block

  12. You can rest your brush softly on the very bottom of your chuck avoiding tissue contact
  13. Pull the brush away easily as the chuck meets the blade

    Catching the curl: as the block meets the blade and
    the section begins its curl, the brush leaves the block
    while catching the curling edge of the section;
    then the brush jumps off the block with the curl

  14. The downward motion of the brush allows you to keep a continuous motion as you take your section

    Pull over the blanket: the brush holding the curl
    pulls the section horizontally over the stage,
    like pulling the blanket over yourself,
    without pressing the tissue to the stage

  15. A glass slide is gently laid upon the tissue section

    Gently touch the section to the slide; avoid stretching or folding the section by keeping a steady hand, and keep the transverse axis of the slide parallel to the section

  16. The tissue section should melt onto the slide
  17. Prepared slides should immediately go into formal alcohol, 95% alcohol (methanol/ethanol) or formalin while awaiting the stain line; if you delay this step, drying artifact will occur
  18. You can take a deeper level after approximately 20 turns (multiple levels may be needed for breast or prostate biopsies)
  19. Optimal cutting thickness is 4 - 7 microns for sectioning and 20 - 40 microns for trimming
Videos


Brush technique
Additional references