Histology Notes

Cutting Grab assigned blocks set up hot water bath set up ice tray melt edges of blocks (bottleneck; take turns, go at different times) Check microtome, turn on Move body to the back. press digital recess button (upper left button at the bottom, up arrow with line) Align block holder (up and down settings, align with red indicator out and white lines) add blade Trim blocks, place on moistened ice tray Always lock handle Always put blade guard up Trim to face the block, then switch to .3 to make it nice before putting it back on the ice tray. After trimming, clean paraffin away, brush behind blade and everywhere, empty tray water bath 47-50°, run tissue paper through to get rid of bubbles start sectioning at 0.3 from first block (should have cooled and moistened enough by now), section slowly and pull on ribbon upwards. if static and or wrinkly, breathe on it. some wrinkles should spread out on water bath. use forceps or fingers. Use pic to free ribbon from blade and together carefully bring ribbon to water bath. Use pic to choose best section and use slide to cut at the weakest point of ribbon or use pic or your finger after it’s out of the water. label slide, put blade guard up, and use tissue to clean water bath. Troubleshooting No Ribbon if ribbon bundles up the block is not cool enough. also check the opposite side of the blade and microtome block, wax builds up there especially after trimming a lot. Thick and thin usually hard tissue like cervix. rotate block. fresh sharp blade. review NaOH procedure Venetian Blinds vibration, some microtomes vibrate on auto mode. check if everything is tight. GI highly susceptible to aggressive trimming. hard to fix once the inner layer of muscle is removed. attempt rotating block to angle with layers or just stop aggressively trimming When to melt down : avoid melting. Save a section and trim deeper. if features are lost, order CUT slide and explain why. melting affects orientation especially after trimming and cutting deeply. really only melt for clips and sutures.

Embedding

confirm cooling is on confirm wax nozzle is adequate CONFIRM wax chamber is filled enough. otherwise the top of cassettes will be at a different temperature push down. can be a bit aggressive. the goal is to make trimming and sectioning easier. GI can take it. Breast be more gentle. use finger or multiple forceps to push more at once. inked margins are helpful to focus on. after embedding and sealing with cassette on top, lift it up to top up with wax, stop sliding it back. Check for bubbles and check for adequate support ie wax after added to mould. Always choose larger mould. i frequently choose one too small. clean station gutters, wax tray, and move rack to processor room when done (there’s a pile somewhere) 🔵 means Note in case 🔴 means tiny piece ?

histo-unloading