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Microtomy
Lab
| Question | Answer |
|---|---|
| Irregular or skipped sections may be caused by: | Too little/much blade tilt, poorly adjusted chuck, cutting too fast or at inconsistent speeds, old or out of adjustment microtome, loose screws/parts |
| Crooked ribbons may be caused by: | Blocks not trimmed parallel on horizontal sides, irregular knife edge, uneven block temp |
| Sections sticking to knife may be caused by: | Dirty knife - clean w/ xylene, knife angle too small, dull knife |
| Smeared/deformed sections may be caused by: | Dull knife |
| Washboarding/undulations in section may be caused by: | Block not clamped tightly, too much knife tilt, loose screws/parts, old microtome, under/overfilled block not sitting properly, often happens w/ hard tissues (e.g. uterus) |
| Ribbon may not form because: | Cold room - warm knife slightly, too much knife angle, too thick sections, dull knife |
| Sections compressed, wrinkled, or jammed together may be caused by: | Dull knife, paraffin build-up on blade, too little knife tilt, cutting too fast, warm room, warm block |
| Knife rings on upstroke & sections are scratched | Knife angle too great > decr. angle Material too hard > soak in water to soften |
| Sections may be lifted from knife on upstroke because: | Too little knife angle, dull knife, warm room - cool block, soft paraffin |
| Knife may not hold edge because: | Knife angle is too severe |
| Scratches or splits in ribbon may be caused by: | Defect in block (e.g. calcium deposit), defect in knife |
| Fragmented or torn sections may be caused by: | Poor processing or fixation, too hot water bath, treat ribbon more gently |
| Sections flying due to static may be helped by: | Breathing on section/blade, humidifer in room, dryer sheet behind blade |
| Chatter may be caused by: | Dry, over-processed tissue, too much knife tilt, cutting too fast, block holder shaft over-extended - vibration |
| Holes in section may be caused by: | Facing block too roughly, tears tissue, can try to face further & smooth |
| Parched earth artifact caused by: | Too hot bath, using cryo spray to cool blocks, facing too roughly, dry sections |
| Wrinkles in section may be caused by: | Not laying ribbon on bath carefully, bath not warm enough |
| Prevent skin cells in sections by: | Wearing gloves, not sticking fingers in bath |
| Prevent cross-contamination by: | Cleaning tools between blocks, cleaning bath between blocks |
| To help keep sections on slide may use: | Slides coated w/ poly L lysine, aminoalkylsilane, or albumin, (may get background staining artifact), charged/plus slides |
Created by:
CCF
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