BRAIN RESEARCH LABORATORIES Histology « Slides * Boxes » Equipment (617) 965-5544 » Fax: (617) 965-6220 1: www brainresearchlab.com INSTRUCTIONS USE AND CARE OF YOUR MICROTOME KNIFE BEFORE USING YOUR MICROTOME KNIFE A D. Your microtome knife has been coated with an oil mixture to prevent rust and corrosion when not in use. Before using your knife, take a lint-free facial tissue saturated in either zylene, benzene, o acetone to remove the protective oil coating on the knife Use a dry, lint-free, facial tissue to wipe your knifé clean, DO NOT USE GAUZE or any other coarse material, it will destroy the edge of your knife, ‘Your knife has already been stropped and is now ready for immediate use. CARING FOR YOUR MICROTOME KNIFE A B. Keep the edge of your knife clean at all times. Spray or brush any household oil on your knife to prevent rust when not in use. Store your knife in its case to prevent oxidation from occurring. Ifyou are using a lab sharpener, periodically send your knife out to be professionally reconditioned 1. Extremely wide facets result from the continuous use of a lab sharpener, The optimum bevel angle becomes distorted as a result of the facet changes. This diminishes your cutting performance 2. Double facets may develop on your knife's edge, creating distorted sections.HINTS FOR USING YOUR MICROTOME KNIFE A. To maximize the sharpness of your knife's edge, use an old knife to trim your first block, or use the extreme end of your new or reconditioned knife. B. Continuously remove the paraffin, or other embedding medium, on the front and back of your knife to maximize your cutting performance C. Depending on the texture of the tissue you have been cutting, you may be able to use the other side of your knife for extended use. D. Setting your knife at the proper clearance angle will obviously not only enhance your output but it will also extend the life of your knife's edge. . Maintain a sharp edge on your knife. Even the most highly skilled technologists cannot overcome the handicaps created from using a dull or bruised knife. F. Based on the facet/bevel angle created in our reconditioning process, we recommend a clearance angle between 6 and 8 degrees. Nl sek G. When your knife is reaching its dimensional maturity, i.., the width of the knife's surface is less than 7/8 of an inch, proper knife stability is difficult to achieve in the holder. Placing a hexagonal pencil at the base of the knife within the holder will act as a filler and provides it with the support required. ‘Microtomy is an Art enhanced by Skilled Technologists Well Maintained Microtomes Sharp, Quality, Microtome Knives Properly Prepared Materials Excellent ProceduresPROBLEM Ribbon does not Crooked ribbons Sections are irveguar, sipped orthin Sectlons are compressed, wrinkled oF Jrmmed together Sections crumble, specimen may tear MICROTOMY SECTIONING TROUBLE-SHOOTING GUIDE cause, Room temperate too old Pali too hard Kifeangl oo great Sections to thick Dui ite Blocks not wimmed, pavallel Irregular kif edge regular paral. softness ‘fone side of block lock temperature uneven dase Tocationneareat Koi it insticent Knife block hole loose Micotome ou of adjustment Dull kite ri ito tight rife edge gummed win para Coming to fs leolicient dehyiaton ofmateril Insulin clearing seater SOLUTION Wan fe slgily Urelower meting point paral “Tiki es outthinner Sharpen kife Reembod in meld sedge ofbock is allt kif edge Use anther part of kaif embed tisue and sr melted parafin, Relocate miertome or ool black in water sje ie holder toa 68 degree searnce angle ‘Tighten st sees ‘Tighten clamp Chece by antares Sharpen ke ‘Coo block in water Increase it Use lino sve tnd sene to dean (Cu thin sections slowly and evenly Savage rarely posible Retum tse several hanger of prin PROBLEM Sections crumble, specimen may ten cout (cont'd) Split ribbon or Tengtise scratches in ribboo Ke rings on up- stroke and sections areserntched Sections ited from kaife on npstroke Sections stlcleto life Scratching noise uring cating cause, Incomplete infttion with embedding medium Exende ie in| peralin bath of| bath oo hot Clearing id used ‘Nicks in aie “Too much it Kaife edge dirty or Tinton edge bjt to hard for palin Hard panicles in parain Cotas ffom rmereunie cherie Fiation Caleeous er sii partes aif ti to peat Material istoo hard Material co hard for parafin method ‘Not enough knife ai Kife ll Room too wan oe parain oo sot rife ee ity rif tit to Title Kieu Material may be twohasd SOLUTION Inflate with parain and reembed (Check thermestat conzole of parain bath Subsite horofomm fortolvee stead of dylene for clearing Sharpen kif Ure lest so kife will eu not sere Clean bot sides of aie xe celoain embedding Rosie paraffin Washing was inant Decal or seals (Change kn it Soak in water 2 alten “Try eeloiin Increase biti Sharpen knife “Try harder prs, euler room oF el Biosk Clean with slene Ivars it Sharpen baie ‘Try calodin embeddingPROBLEM cause. SOLUTION Undulatonsin —Setscrews loose Tiguan set screns the surface ofthe section Krife older loose Tighten lamp Excessive knife it Decrease knife tik Sections are Dall nite Sharpen knife smeared, deformed ROBLEM cause SOLUTION Section fal out of | Enbedding medium —Changeto different rmatris oF show inadequate medium for ‘compression different embedsing from embedding media frame Sections have Inslicient Salvage rarely ‘mushy appearance dehyation possible Kolfe doesnot hold Angletoossvere Adjust clearance Sections My and Static eletrity Increase humidity ies edge angle to 68 degrees siketo parts of Imicrotome or Bil water in a pan Cutingtoofest Slow cutting sped ‘other nearby objects Ground microtome Paralfin oohard Alter consistency of toavate pipe parla Tonize air CCutinghard issue No selution, natural HELPFUL SECTIONING TIPS Section Quality Cutting Speed Seaton quality i determined mor by the ‘ondtion ofthe knife andthe clearance ange, than by any oie factors Clearance Angle ‘Based onthe bevel angle rete in th mars- facturing and reconditioning proce, clearance angle betwen 6 and 8 dgres is recommended “Maximizing Cutting Performance Maintain a sharp edge on your kif. Even the ‘mod highly aed technologie cannot overcome the hmdcape created fom using a dul cr baie knife Koife Cleanliness CContnsously remove the paraffin or other embedding medi from he nt andthe back of your ke, {o achiev optimum section quali. “The cutting spend for etng ood parafin sections iearate at which the mirotome deve wheel eater ppronimatlyone revolution per second. Dimensional Maturity ‘When the width ofthe kif surface i as that 7/8 ‘ofan inch prope a ability sfc to achieve and the ie shouldbe replaced. Kafe Support ‘When your knife ie approaching its dimensional ratty, lacing «hexagonal pencil atthe base ofthe holder will ‘stars filler nd provide your knife withthe support eqired for sabi. Reversing Cuting Edge Use the oppasite side of your kati section quality is sot acepable, This procedure may aso extnd the use- Tite of your kif, Microtomy is an Art Skilled Tecknologits Sharp, Quality Microtome Knives Well Maintained Microtomes Properly Prepared Meteriais Excellent Procedures