Title Leica RM2125 RTS & Olympus microTec CUT 4060 Written Instructions School of Medicine Faculty Health Prepared By Monique Trengove on 14/06/2017 Megan Ellis Authorised By Elizabeth Laidlaw – Senior Technical Officer on 19/6/2017 Reviewed By Elizabeth Laidlaw On 19/6/2017
Microtome
Basic operation and safety procedures The following document identifies the instructions and safety procedures required when using School of Medicine microtomes - Leica RM2125 RTS (ka2.225) & Olympus microTec CUT 4060 (nb1.115) microtomes. **All users must be trained prior to using SOM microtomes
Description of Leica RM2125 RTS & Olympus microTec CUT 4060 microtomes: Microtomes are used to cut thin slices of material, known as sections. Microtomes are used in microscopy, allowing for the preparation of samples for observation under transmitted light or electron radiation.
Users must supply the following: • Samples mounted in paraffin wax on histological cassettes • Knives/blades
SOMHisto will supply the following: • Paint brushes • Forceps
Preparation of samples: Please consult appropriate literature for preparation of your samples. Tissue processor and mounting equipment available in nb1.115 (training required).
Simple Instructions:
1. Clamping the specimen Always clamp the specimen block BEFORE clamping the knife or the blade.
• Rotate the handwheel (RHS) until the specimen clamp is in the uppermost position. • Activate the handwheel lock. • To insert a specimen into the specimen holder, push the lever forwards. • Mount the cassette horizontally or vertically as required.
Leica RM2125RTS Olympus microTec CUT
Specimen Specimen holder holder Specimen lever holder lever Specimen holder
2. Clamping the knife / disposable blade
Take care when handling microtome knives and disposable blades. The cutting edge is extremely sharp and can cause serious injuries!
• Lower the knife guard on the knife holder. • Rotate the clamping lever downwards (unclamped) • Carefully insert the blade from the side • With the blade in position, rotate the clamping lever upwards (clamped) • Once the clade is securely clamped, raise the knife guard into position.
Leica RM2125RTS Olympus microTec CUT
Knife Clamping Knife Clamping lever guard lever guard
3. Adjusting the clearance angle
• Procedure specific to microtome model (refer to relevant manual).
4. Orienting the specimen • Specimen orientation allows for simple position correction of the specimen surface when the specimen is clamped into place. • Run the specimen to the rear end position by turning the coarse feed wheel (left hand side) • Loosen the clamping lever in front on the microtome base plate and slide the knife holder base with knife holder until it is almost in front of the specimen.
Handwheel
Knife holder
Knife holder clamping lever
• Move the specimen holder to the upper end position by turning the handwheel and engaging the handwheel lock. • Release the specimen holder clamp, turn the eccentric lever counterclockwise. • Turn setscrew #1 to orient the specimen in north-south direction. • Turn setscrew #2 to orient the specimen in east-west direction. • To lock the orientation, turn the eccentric lever clockwise.
Leica RM2125RTS Olympus microTec CUT
Eccentric Setscrew Setscrew lever #1 #1 Eccentric lever
Setscrew Setscrew #2 #2
5. Trimming the specimen • Use the coarse feed wheel (left hand side) for a fast horizontal forwards movement of the specimen – towards the knife – and backwards – away from the knife. • Once in position, set to a large section thickness (eg. 50uM). • Bring the specimen closer to the knife by rotating the coarse feed wheel and trim it by simultaneously rotating the handwheel (right hand side). • Trim until the desired specimen plane is reached (procedures may vary between microtome models, please see appropriate manual for detailed instruction).
6. Sectioning • Set the desired section thickness using the thickness setting knob at the front of the microtome on the right and check the value in the display window.
Leica RM2125RTS Olympus microTec CUT
Section Section Thickness thickness thickness setting knob Thickness setting knob
• Use a different area of the cutting edge (blade) for trimming than for sectioning. **Use caution when repositioning blades and always ensure they are clamped back in place. • For sectioning, turn the handwheel (right hand side) evenly in a clockwise direction. Repeat until desired number of sections are cut. • Pick up the sections and mount them on microscope slides by preferred method.
**Always rotate the handwheel at a uniform speed. The rotation speed of the handwheel must be adapted to suit the hardness of the specimen. For harder specimens, use a slower speed.
7. Changing specimens • Cover the sectioning edge with the knife guard.
• Lock the handwheel • Remove the specimen from the specimen clamp and mount a new specimen. • Repeat step 2-4 to position and prepare the new sample.
8. Pack up • Move the specimen clamp to the upper end position and activate the handwheel lock.
• Remove the blade from the knife holder and insert it in the receptacle at the bottom of the dispenser, or remove the knife from the knife holder and put it back in the knife case.
• Remove knife holder base and knife holder for cleaning.
• Remove the specimen from the specimen clamp.
• Remove section waste with a dry paint brush.
• Remove specimen clamp and clean separately.
Cleaning If necessary, the varnished outside surfaces can be cleaned with a mild commercial household cleaner or soap water and then be dried with a moist cloth. Do not use any of the following for cleaning the outside surfaces of the instrument: alcohol, detergents containing alcohol (window cleaner!), abrasive cleaning powders, solvents containing acetone or xylene. Xylene or acetone will damage the finished surfaces!
Safety Precautions
***Lock the handwheel (right hand ride) and cover the knife edge with the knife guard prior to any manipulation of knife or specimen, prior to changing the specimen block and during all work breaks!***
Always remove the knife / blade before detaching the knife holder from the instrument.
Always put the knives back into the knife case when not in use!
Never place a knife anywhere with the cutting edge facing upwards and never try to catch a falling knife!
SOMHisto SOM Histology Facility Link Paraffin Embedded Tissue Cutting link