RM2135 Instruction Manual Ver 1.1 Feb 2001

NOTE The information, numerical data, notes and value judgments contained in this manual represent the current state of scientific knowledge and state-of-the-art technology as we understand it following thorough investigation in this field. We are under no obligation to update the present manual periodically and on an ongoing basis according to the latest technical developments, nor to provide our customers with additional copies, updates etc. of this manual. For erroneous statements, drawings, technical illustrations etc. contained in this manual we exclude liability as far as permissible according to the national legal system applicable in each individual case. In particular, no liability whatsoever is accepted for any financial loss or consequential damage caused by or related to compliance with statements or other information in this manual. Statements, drawings, illustrations and other information as regards contents or technical details of the present manual are not to be considered as warranted characteristics of our products.

These are determined only by the contract provisions agreed between ourselves and our customers. Leica reserves the right to change technical specifications as well as manufacturing processes without prior notice. Only in this way is it possible to continuously improve the technology and manufacturing techniques used in our products. This document is protected under copyright laws. Any copyrights of this document are retained by Leica Microsystems Nussloch GmbH. Any reproduction of text and illustrations (or of any parts thereof) by means of print, photocopy, microfiche, web cam or other methods – including any electronic systems and media – requires express prior permission in writing by Leica Microsystems Nussloch GmbH. For the instrument serial number and year of manufacture, please refer to the name plate at the back of the instrument.

© Leica Microsystems Nussloch GmbH

Published by: Leica Microsystems Nussloch GmbH Heidelberger Str. 17 - 19 D-69226 Nussloch Germany Phone: +49 (0)6224 143-0 Fax: +49 (0)6224 143-200 Internet: http://www.histo-solutions.com Leica RM2235

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Table of contents 1. Important notes ... 6 2. Safety ... 7 2.1 2.2 2.3

Safety notes ... 7 Warnings ... 7 Integrated safety devices ... 10

3. Instrument components and specifications ... 12 3.1 3.2 3.3

Overview - instrument components ... 12 Instrument specifications ... 13 Technical data ... 14

4. Startup ... 15 4.1 4.2 4.3 4.4 4.5

Standard delivery ... 15 Installation site requirements ... 15 Assembling the handwheel ... 16 Inserting the universal cassette clamp ... 17 Inserting the knife holder ... 18

5. Operation ... 19 5.1 Operating elements and their functions ... 19 5.1.1 Section thickness setting ... 19 5.1.2 Coarse driving wheel ... 19 5.1.4 Mechanical trimming function ... 20 5.1.3 Specimen retraction ... 20 5.1.5 Specimen holder with precision orientation ... 21 5.2 Adjusting the clearance angle ... 22 5.3 Fine adjustment of the mass balance ... 23 5.4 Clamping the specimen ... 24 5.5 Clamping the knife / disposable blade ... 24 5.6 Sectioning ... 25 5.7 Changing the specimen or interrupting sectioning ... 26 5.8 Finishing daily routine ... 26

6. Optional accessories ... 27 6.1 Installing the fixture for specimen clamps ... 27 6.1.1 Non-orienting fixture f. spec. clamps ... 27 6.1.2 Orienting fixture f. specimen clamps ... 27 6.1.3 Fixture for specimen clamps with precision orientation ... 28 6.1.4 Quick-clamping system ... 29

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Instruction manual V 1.1 – 02/2005

Table of contents 6.2 Specimen clamps and holders ... 30 6.2.1 Standard specimen clamp ... 30 6.2.2 Vee insert ... 30 6.2.3 Foil clamp type 1 ... 31 6.2.4 Foil clamp type 2 ... 32 6.2.5 Universal cassette clamp ... 32 6.2.6 Super Mega cassette cla mp ... 33 6.2.7 Holder for round specimens ... 34 6.3 Knife holder base and knife holder ... 35 6.3.1 Knife holder base, without lateral movement feature ... 35 6.3.2 Knife holder E/E-TC ... 36 6.3.3 Knife holder N/NZ ... 38 6.4 Section waste tray ... 40 6.5 Backlighting ... 40 6.7 Freezer pack ... 41 6.6 Tray ... 41 6.8 Ordering information ... 42

7. Trouble shooting ... 44 7.1 Possible faults ... 44 7.2 Instrument malfunctions ... 44

8. Cleaning and maintenance ... 45 8.1 8.2 8.3

Cleaning the instrument ... 45 Maintenance instructions ... 47 Lubricating the instrument ... 48

9. Warranty and service ... 49

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1. Important notes Symbols in the text and their meanings Dangers, warnings and cautions appear in a gray box and are marked by a warning triangle .

Notes, i.e. important user information appear in a gray box and are marked by an information symbol .

(5)

Intended use of instrument The Leica RM 2235 is a manually operated rotation microtome for creating thin sections of specimens of varying hardness for use in routine and research laboratories in the fields of biology, medicine and industry. They are designed for cutting soft paraffin specimens as well as harder specimens, as long as they are suitable for being cut manually.

Any other use of the instrument is considered improper!

Numbers in parentheses refer to item numbers in illustrations.

Qualification of personnel

Instrument type

• The Leica RM 2235 may be operated by trained laboratory personnel only.

All information provided in this manual applies only to the instrument type indicated on the cover page. An identification label indicating the instrument serial number is attached at the left side of the instrument.

• All laboratory personnel designated to operate the Leica instrument must read this instruction manual carefully and must be familiar with all technical features of the instrument before attempting to operate it.

Fig. 1

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Instruction manual V 1.1 – 02/2005

2.

Safety

Be sure to comply with the safety instructions and warnings provided in this chapter. Be sure to read these instructions, even if you are already familiar with the operation and use of other Leica products.

2.1

Safety notes

This instruction manual contains important instructions and information regarding the operational safety and maintenance of the instrument. The instruction manual is an important part of the product, which must be read carefully prior to startup and use and must always be kept near the instrument. If additional requirements on accident prevention and environmental protection exist in the country of operation, this instruction manual must be supplemented by appropriate instructions to ensure compliance with such requirements.

This device has been built and tested in accordance with the directive for machines, 98/37/EC, and the safety regulations governing laboratory apparatus. The following standards serve as a basis: • • • •

DIN EN ISO 12100-1: 2003 DIN EN ISO 12100-2: 2003 DIN EN 61010-2-101: 2002 DIN EN ISO 9001: 2000.

In order to maintain this condition and ensure safe operation, the operator must observe all the instructions and warnings contained in this instruction manual.

The protective devices on both instrument and accessories may neither be removed nor modified. Only service personnel qualified by Leica may repair the instrument and access the instrument’s internal components.

2.2 Warnings The safety devices installed in this instrument by the manufacturer only constitute the basis for accident prevention. Primarily responsible for accident-free operation is above all the owner of the instrument and, in addition, the designated personnel who operates, services or cleans the instrument. To ensure trouble-free operation of the instrument, make sure to comply with the following instructions and warnings. Leica RM2235

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2. Safety Warnings - Safety instructions / warning labels attached to the instrument • Safety instruction labels on the instrument marked with a warning triangle indicate that the correct operating instructions (as described in this manual) must be followed when operating or replacing the instrument component bearing the label. Failure to adhere to these instructions may result in an accident, personal injury, damage to the instrument or accessory equipment.

Warnings - Transport and installation • Once removed from the crate, the instrument may only be transported in an upright position. • Do not transport the instrument by holding it by the handwheel grips, coarse driving wheel or the knob for setting the section thickness. • The protective devices on both instrument and accessories must neither be removed nor modified.

Warnings - Working at the instrument

• Take care when handling microtome knives and disposable blades. The cutting edge is extremely sharp and can cause severe injury! • 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! • Always clamp the specimen block BEFORE clamping the knife.

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Instruction manual V 1.1 – 02/2005

2.

Safety

Warnings - Working at the instrument • Prior to manipulating the knife and specimen, or changing the specimen or knife, and during breaks, always lock the handwheel and cover the cutting edge with the knife guard! • ALWAYS rotate the handwheel clockwise. Otherwise, the locking mechanism will not work properly. • Always wear protective glasses when sectioning brittle specimens. Specimen may splinter! • Specimen blocks must not be oriented during the retraction phase! If a block is oriented during retraction, the block will advance by the retraction value PLUS the selected section thickness before the next section. This may cause damage to both specimen and knife!

Warnings – Cleaning and maintenance • Always lock the handwheel before cleaning! • Do not use any solvents containing acetone or xylene for cleaning! • Ensure that no liquids enter the interior of the instrument when cleaning! • When using detergents please comply with the safety precautions of the manufacturer and with the applicable laboratory regulations.

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2. Safety 2.3 Integrated safety devices Locking the handwheel There are two ways of locking the handwheel (12): Using the lever (3) on the right side of the microtome base plate, the handwheel can be braked in almost any position. • To brake, rotate the lever in a counterclockwise direction to position
.

5

Caution! The braking lever (3) must be exactly in position
, so that the handwheel brake is applied correctly. If the lever is moved beyond this point, it is possible that the handwheel is no longer braked.

12

• To unlock the handwheel brake turn the lever (3) back to its original position. Position

. • To lock the handwheel, press the lever (5) outwards and continue to turn the handwheel slowly until it locks exactly in the 12 o’clock position.

3

In either case, the yellow LED in the LOCK field (4) lights up and the instrument cannot be started.

Fig. 2

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If both braking systems are used simultaneously, always bring the lever (3) into position  first to release the brakes. Otherwise it is possible that the lever (5) cannot be unlocked.

Instruction manual V 1.1 – 02/2005

2. Safety Knife guard on the knife holder

Knife holder N

7

8

7

Each knife holder is equipped with a tightly mounted knife guard (8, 9). This makes it possible to cover completely the cutting edge in every knife or blade position. Knife holder N/NZ

Fig. 3

Knife holder E/E-TC

Knife holder E 11

9

The knife guard (8) of the knife holder N/NZ can be easily positioned via the two buttons (7) (Fig. 3). To cover the knife edge, push both cover strips of the knife guard to the center.

The knife guard on knife holder E/E-TC consists of a red foldaway handle (9). To cover the cutting edge, fold the knife guard handle (9) upwards as illustrated in Fig. 4.

10

The clamping levers on the knife holder E are not interchangeable. The two clamping levers (10, 11) must remain in the position shown at all times, as otherwise isolated malfunctions of the knife holder can occur. Clamping lever for the blade (10) at the right, clamping lever for the lateral displacement (11) at the left.

Knife holder E-TC

9 Fig. 4

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3.

Instrument components and specifications

3.1 Overview - instrument components Tray

Universal cassette clamp

Handwheel locking mechanism

Knife holder base, without lateral movement feature

Smooth-turning handwheel

Lever for activating the handwheel brake

Removable section waste tray

Clamping lever of the knife holder base Window for displaying the section thickness

Directional fixture for specimen clamps

Adjusting knob for setting the section thickness

Coarse driving wheel

Lever for activating the mechanical trimming function

Clamping lever for lateral movement function of knife holder

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Knife holder E

Fig. 5

Instruction manual V 1.1 – 02/2005

3. Instrument components and specifications 3.2 Instrument specifications Basic Instrument with mechanical trimming, with lateral coarse feed (clockwise (CW) or counter clockwise (CCW)) without specimen orientation The Leica RM2235 rotary microtome features low-maintenance and backlash-free precision micrometer feed system with horizontal feed and vertical stroke mechanism based on self-lubricating cross roller bearings. For increased user safety, the instrument isequipped with two independent handwheel lockingsystems. The instrument is equipped with a Leica-patented operator-adjustable force balancing system with spring compensation of the forces which occurduring the sectioning process. Advantages: no need of a heavy counter balance in the handwheel and possibility to adapt the spring force to specimen clamps and specimens of different weights. A patented specimen retraction system can be switched ON/OFF by the operator, a feature, which provides the full advantage of an instrument with specimen retraction while at the same time making it easy to operate the instrument in the ‘rocking mode’ (if the in strument is used in the ‘rocking mode’, we recommend to switch the specimen retraction OFF). The coarse feed wheel is within easy reach of the operator. (more information to direction of rotation see chapter 5.1.2)

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3.

Instrument components and specifications

3.3 Technical data General Approvals: Operating temperature range: Storage temperature range: Relative humidity: Humidity in storage loaction: Section thickness range:

The instrument-specific marks are located on the rear panel of the instrument next to the name plate +10 °C to +35 °C +5 °C to +55 °C max. 80%, non-condensing < 80 % 1.0 - 60.0 µm

Section thickness settings :

from 1.0 - 10.0 µm in 1.0 µm increments from 10.0 - 20.0 µm in 2.0 µm increments from 20.0 - 60.0 µm in 5.0 µm increments

Specimen feed: Vertical stroke: Maximum sectioning area w/o retraction: Maximum sectioning area with retraction: The specimen retraction:

approx. 30 mm 70 mm 69 mm (without specimen orientation 1 µm) 62 mm 200 µm; can be turned off manually.

Dimensions and weight Width (including handwheel): Depth (including waste tray): Height (total): Working height (knife blade): Weight (without accessories)

413 mm 618 mm 305 mm (with tray on the housing) 168 mm (measured from the table) approx. 37 kg

Optional equipment and optional accessories Specimen orientation (option) horizontal: 8° vertical: 8° Angle of rotation: ± 90°

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Trimming stages :

10 µm, 50 µm

Repositioning of knife holder base north-south:

± 25 mm Instruction manual V 1.1 – 02/2005

4. Startup 4.1 Standard delivery The Leica RM 2235 standard delivery includes: 1 Leica RM2235 basic instrument 1 Handwheel, complete 1 Section waste tray 1 Tool set consisting of: 1 Allen key with handle, size 5 1 Allen key with handle, size 4 1 Allen key size 3 1 Screwdriver 3 x 50, 186 long 1 Bottle (50 ml) of oil for drives, type 405 1 Dust protective cover 1 Brush with magnet 1 Operating manual G/E/F/S The accessories ordered are included in a separate box. Carefully check the delivery against the packing list and the delivery note. Should there be any discrepancy, please contact the Leica selling unit handling your order.

0500 38181 0502 37931 0500 38600 0194 04760 0194 04782 0222 04138 0170 11568 0336 06086 0212 30350 0183 40426 0700 37115

4.2 Installation site requirements • Stable, vibration-free laboratory bench with horizontal and even stage plate; practically vibration-free floor. • No other instruments nearby which might cause vibrations. • Room temperature permanently between + 10 °C and + 35°C. • Obstruction-free access to the handwheel.

Never operate the instrument in rooms with explosion hazard.

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4.

Startup

Setting up the instrument 5

Do not transport the instrument by holding it on the handwheel shaft, the object head or the housing! 6

• By holding the instrument (5) at the front by the baseplate, and at the rear by the recessed grip (Fig. 6), lift it out of the molded cushion (6) of the packaging and place it on a stable laboratory table. • The two sliding elements (9) located on the rear of the base plate make it easier to move the instrument on the table.

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Fig. 6

• To move the instrument, hold it by the front of the base plate, lift it up gently and slide it on its slides.

4.3 Assembling the handwheel The handwheel has to be assembled before attempting to use the instrument. The necessary parts and tools can be found in the toolkit supplied together with the instrument. 4 3

1

2

2a

Abb. 7

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The feather key (4) is loosely placed in the handwheel shaft (1) and fixed in place with a tie-rap during transport. • Remove the cable tie (3). Caution! Make sure not to lose the feather key! • Place the handwheel (2) on the handwheel shaft (1) as shown. • Tighten the screw (2a) located in the center hole of the handwheel with an Allen key size 4 (5). • Remove the cover foil from the self-adhesive cover disk and fix the cover disk on the handwheel.

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Instruction manual V 1.1 – 02/2005

4. Startup 4.4 Inserting the universal cassette clamp There are two versions of the specimen holder, one with and one without specimen orientation, which are interchangeable. The object orientation allows for simple position correction of the specimen surface when the specimen is clamped into place.

60

You can use the quick clamping system (64) to hold all available accessory specimen clamps (for more information, see Chapter 6 “Optional accessories“).

12

To do so, proceed as follows: • Move the object head (60) to the upper end position by turning the handwheel (12) and engage the handwheel lock. 60

62

64

• To release the clamping system, turn the screw (61) of the quick clamping system (64) counterclockwise using an Allen key size 4 (71). • Push the guide (63) of the universal cassette clamp (62) from the left into the quick clamping system (64) as far as it will go.

61 71

63

• To clamp the cassette clamp turn the screw (61) clockwise as far as it will go.

Fig. 8

Since all stage clamps available as accessories are equipped with the same kind of guide on the back, they are inserted in the same way described here using the example of the cassette clamp.

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4. Startup Setting up the knife holder base

4.5 Inserting the knife holder

• Release the clamping lever (50) by rotating it counterclockwise. • Insert the knife holder base (51) using the notch (52) on the bottom into the T-piece (55) of the microtome base plate (53). • To secure the knife holder base, turn the clamping lever (50) clockwise.

55 53

The knife holder base (51) can be moved back and forth on the microtome base plate. This allows bringing the knife holder to optimal cutting position in relation to the specimen. There is a scale (54) on the right side of the microtome base plate. This enables faster and better positioning of the knife holder at the specimen if various combinations of standard specimens and specimen holders are used. The rear edge of the knife holder base (51) functions as the scale reference.

50 52

Enlarged detail: Scale for better repositioning of the knife holder for varying specimen heights.

51

54 Fig. 9

56

71

Inserting the knife holder • Release the screw (58) with the size 4 Allen key (71) (part of the delivery package) so that the knife holder (57) can be moved. • Place the knife holder (57) with the underside groove onto the T-piece (56) of the knife holder base (51).

57

• To clamp, retighten the screw (58). 58 Fig. 10

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Instruction manual V 1.1 – 02/2005

5. Operation 5.1 Operating elements and their functions

The section thickness is set by turning the adjusting knob (33) at the front of the microtome on the right. The adjusting knob has a notch for each value that can be set. Setting range: 1 - 60 µm from 1 - 10 µm in 1 µm increments from 10 - 20 µm in 2 µm increments from 20 - 60 µm in 5 µm increments.

38

33

5.1.1 Section thickness setting

34

The section thickness set in each case is displayed in the window (34). The selected section thickness (on the scale) must agree with the red pointer (38)

5.1.2 Coarse driving wheel Fig. 11

The instrument can be ordered with clockwise or counterclockwise rotation. The given direction of rotation means “forwards” and relates to the feed movement of the specimen towards the knife. The coarse motion serves for a fast horizontal forwards movement of the object - towards the knife - and backwards - away from the knife. When reaching the rear/front end positions, the coarse driving wheel can only be turned with difficulty. In the front end position, no more feed motion takes place. The coarse driving wheel also turns during sectioning. Therefore it must not block whilst the handwheel is being turned during sectioning; otherwise, no feed motion can take place and thus also no sectioning.

Fig. 12

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5. Operation 5.1.3 Specimen retraction Magnet for holding the wrench size 4

For turning the retraction Off, turn the slot into the horizontal position (off). For turning the retraction On, turn the slot into the vertical position (on).

The specimen retraction serves for protecting the knife and the specimen. When the retraction is switched on, the object is drawn back 200 µm into the upper end position after the sectioning stroke during the return movement. Before the feed motion of the new section thickness, the feed motion for the retraction value takes place. The specimen retraction can be switched off manually at the back of the instrument (Fig. 13), if required, using the slotted-screwdriver supplied with the delivery. Before switching the specimen retraction On and Off, run the object head to the upper end position by turning the handwheel.

Fig. 13

5.1.4 Mechanical trimming function The RM 2235 is fitted with a mechanical trimming function. The trimming lever has 3 notching positions (0,10 µm, 30 µm). The points (36) mark the two trimming stages:
= 10 µm

= 30 µm 36

Fig. 14

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• For activating the trimming function, press the lever downwards into one of the two notching positions and keep depressed. After each rotation of the handwheel, a feed motion of 10 µm or 30 µm takes place. • After letting go of the lever, it automatically springs back to its original position (zero position). The trimming function is thereby deactivated. The section thickness that has been set is not added to the selected trimming value. If the section thickness that has been set is greater than the selected trimming value, the section thickness is fed. Instruction manual V 1.1 – 02/2005

5. Operation 5.1.5 Specimen holder with precision orientation In the quick clamping device of the specimen holder fixture with precision orientation, all object clamps available as optional accessories can be used (implemented).

30

The object orientation allows for simple position correction of the specimen surface when the specimen is clamped into place.

32

32

Orienting the specimen Specimen blocks must not be oriented during the retraction phase! If a block is oriented during retraction, the block will advance by the retraction value PLUS the selected section thickness before the next section. This may cause damage to both specimen and knife!

31 29

Fig. 15

Display of the zero position For better display of the zero position, the orientation has two red indicators (32). When both indicators are visible and both setscrews are in zero position at the same time (notch point, white marking on “
“), the specimen is in zero position. When the large standard clamp (50 x 55 mm) is used, the specimen orientation of 8° in north-south direction is no longer possible. The usable angle is only about 4° in this case.

Leica RM2235

• Raise the object head to the upper end position and activate the handwheel lock. • To release the clamp, turn the eccentric lever (29) forwards. • Turn setscrew (30) to orient the specimen in north-south direction. Turn setscrew (31) to orient the specimen in east-west direction. Each complete turn of the screw inclines the specimen by 2°. A total of 4 complete turns = 8° are possible in every direction. The accuracy is approximately ± 0.5°. For ease of estimation, there is a white marking on the handle and a notch point that is noticeable during turning. • To lock the current orientation, turn the eccentric lever (29) backwards.

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