RM2235 Instruction for Use Ver 1.8 Rev C Sept 2013

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. To the extent permitted in accordance with the national legal system as applicable in each individual case, we shall not be held liable for erroneous statements, drawings, technical illustrations etc. contained in this manual. 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 Instructions for Use 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. All copyrights to this documentation are held by Leica Biosystems 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 Biosystems Nussloch GmbH. For the instrument serial number and year of manufacture, please refer to the nameplate on the back of the instrument. © Leica Biosystems Nussloch GmbH

Leica Biosystems Nussloch GmbH Heidelberger Str. 17 - 19 D-69226 Nussloch Germany Phone: +49 62 24 143-0 Fax: +49 6224 143-268 Internet: http://www.LeicaBiosystems.com Assembly contracted to Leica Microsystems Ltd. Shanghai Leica RM2235

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Table of Contents 1. 2.

Important Information... 4 Safety... 5 2.1 2.2 2.3

3.

Instrument Components and Specifications... 10 3.1 3.2 3.3

4.

Standard delivery... 13 Installation site requirements... 13 Unpacking and installation... 14 Assembling the handwheel... 16 Inserting the universal cassette clamp... 17 Inserting the knife holder... 18

Operation... 19 5.1 5.1.1 5.1.2 5.1.3 5.1.4 5.1.5 5.2 5.3 5.4 5.5 5.6 5.7 5.8

6.

Overview - instrument components... 10 Instrument specifications... 11 Technical Data... 12

Instrument Setup... 13 4.1 4.2 4.3 4.4 4.5 4.6

5.

Safety notes... 5 Warnings... 5 Integrated safety devices... 8

Operating elements and their functions... 19 Section thickness setting... 19 Coarse driving wheel... 19 Specimen retraction... 20 Mechanical trimming function... 20 Specimen holder with precision orientation... 21 Adjusting the clearance angle... 22 Fine adjustment of the force balance... 23 Clamping the specimen... 24 Clamping the knife / disposable blade... 24 Sectioning... 25 Changing the specimen or interrupting sectioning... 26 Finishing the daily routine... 26

Optional Accessories... 27 6.1 Assembly for fixture for specimen clamps... 27 6.1.1 Rigid fixture for specimen clamps... 27 6.1.2 Directional fixture for specimen clamps... 27

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Instructions for Use V 1.8 RevC – 09/2013

Table of Contents 6.1.3 Fine-directional fixture for specimen clamps... 28 6.1.4 Quick clamping system... 29 6.2 Specimen clamps and holders... 30 6.2.1 Standard specimen clamp... 30 6.2.2 Vee insert... 31 6.2.3 Foil clamp type 1... 32 6.2.4 Universal cassette clamp... 33 6.2.5 Super mega-cassette clamp... 34 6.2.6 Holder for round specimens... 35 6.3 Knife holder base and knife holder... 36 6.3.1 Knife holder base, without lateral displacement feature... 36 6.3.2 Knife holder E/E-TC... 37 6.3.3 Knife holder N/NZ... 40 6.4 Section waste tray... 42 6.5 Backlighting... 42 6.6 Tray... 43 6.7 Freezer pack... 43 6.8 Universal microscope carrier ... 44 6.9 Magnifying lens ... 46 6.10 Additional accessories ... 47 6.11 Ordering information... 62

7.

Troubleshooting... 64 7.1 7.2

7. 8.

Troubleshooting... 66 Cleaning and Maintenance... 67 8.1 8.2 8.3

9. 10.

Possible faults... 64 Instrument malfunctions... 66

Cleaning the instrument... 67 Maintenance instructions... 69 Lubricating the instrument... 70

Warranty and Service... 71 Decontamination Certificate (Master)... 72

Leica RM2235

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1.

Important Information

Symbols in the text and their meanings Warnings appear in a gray box and are marked by a warning triangle .

the Leica instrument must read these Instructions for Use carefully and must be familiar with all technical features of the instrument before attempting to operate it.

Designated use Notes, i.e. important user information appears in a gray box and is marked by an in. formation symbol

(5)

Numbers in parentheses refer to item numbers in illustrations.

Manufacturer Date of Manufacture This product fulfills the requirements of the Council's Directive 98/79/EC concerning in vitro diagnostics (IVD) medical devices. In vitro diagnostics (IVD) medical ­device Observe the Instructions for Use

The Leica RM2235 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. It is designed for sectioning soft paraffin specimens as well as harder specimens, as long as they are suitable for being cut manually. Any other use of the instrument will be considered as improper use!

Instrument type All information provided in these Instructions for Use applies only to the instrument type indicated on the title page. A nameplate indicating the instrument serial number is attached at the left side of the instrument. Fig. 1 is provided as an example only and shows a valid nameplate for this instrument.

Order No. Serial number

Qualification of personnel • The Leica RM2235 may be operated by trained laboratory personnel only. • All laboratory personnel designated to operate

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Fig. 1 Instructions for Use V 1.8 RevC – 09/2013

2.

Safety

The safety and caution notes in this chapter must be observed at all times. Be sure to read these notes even if you are already familiar with the operation and use of other Leica products.

2.1

Safety notes

These Instructions for Use include important information related to the operating safety and maintenance of the instrument. The Instructions for Use are an important part of the product, and must be read carefully prior to startup and use and must always be kept near the instrument. These Instructions for Use must be appropriately supplemented as required by the existing regulations on accident prevention and environmental safety in the operator‘s country.

This instrument is built and inspected according to the Safety requirements for laboratory instruments. The current EC Declarations of Conformity can be found on the Internet: www.LeicaBiosystems.com To maintain this condition and ensure safe operation, the user must observe all notes and warnings contained in these Instructions for Use.

The protective devices located on the instrument and the accessories must not be removed or 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. Operating the instrument safely is, above all, the responsibility of the owner, as well as the designated personnel who operate, service or clean 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 notes on the instrument itself • Safety notes on the instrument itself, which are marked with a warning triangle, indicate that the correct operating instructions (as defined in these Instructions for Use) must be followed when operating or replacing the item marked. • 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 unpacked, the instrument may be transported only 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 located on the instrument and the accessories must not be removed or modified.

Safety instructions - working with the instrument • Take care when handling microtome knives and disposable blades. The cutting edge is ­extremely sharp and can cause serious injuries! • 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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Instructions for Use V 1.8 RevC – 09/2013

2.

Safety

Safety instructions - working with 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 turn the handwheel clockwise; otherwise, the brake will not work properly. • Always wear safety goggles when sectioning brittle specimens. Specimens 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! • Prior to sectioning, check that the specimen is securely clamped in the specimen clamp – failure to observe this poses the risk of damaging the specimen.

Hazards - servicing and cleaning • Always lock the handwheel before cleaning! • Do not use any solvents containing acetone or xylene for cleaning! • Ensure that liquids do not enter the interior of the instrument during cleaning! • When using cleaners, please comply with the safety instructions of the manufacturer and the laboratory safety regulations!

Leica RM2235

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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

When using both brake systems at the same time, always move the lever (3) to position  first. Otherwise, it may be not be possible to release the lever (5). Fig. 2

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Instructions for Use V 1.8 RevC – 09/2013

2. Knife guard on the knife holder

Knife holder N

7

Safety

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

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9

The knife guard (8) of the knife holder N/NZ can be easily positioned via the two handles (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

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Fig. 4 Leica RM2235

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3.

Instrument Components and Specifications

3.1

Overview - instrument components

Tray

Handwheel locking mechanism

Universal cassette clamp

Knife holder base, non-orientable

Removable section waste tray

Smoothturning Handwheel Lever for activating the Handwheel brake Clamping lever of the knife holder base Window for displaying the section thickness

Directional specimen holder fixture

Adjusting knob for setting the section thickness

Coarse feed wheel

Lever for activating the mechanical trimming function Clamping lever for lateral displacement

10

Knife holder E

Fig. 5 Instructions for Use V 1.8 RevC – 09/2013

3. 3.2

Instrument Components and Specifications

Instrument specifications Basic instrument with mechanical trim function, lateral coarse drive The Leica RM2235 rotary microtome is equipped with a low-maintenance, slack-free micrometer drive, with vertical and horizontal specimen feed realized via low-maintenance cross roller bearings. The instrument is equipped with two independent handwheel locking systems for even greater operating safety. Leica's patented, user-adjustable force balance system compensates centrifugal forces arising while cutting via a pretensioned spring for extremely light handwheel action. Advantage: a heavy counterweight in the handwheel is no longer needed. The spring tension is individually adjustable, corresponding to the weight of the respective attached specimen clamp or specimen. The patented specimen retraction system can be switched on and off by the user. The instrument thus offers all of the advantages of specimen retraction while supporting work in "rocking mode", i.e. without a full handwheel rotation. We recommend disabling specimen retraction when working in "rocking mode". The coarse drive wheel is ergonomically positioned. (For more information on the rotating direction, see chapter 5.1.2)

Leica RM2235

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3.

Instrument components and specifications

3.3

Technical Data

General Approvals: The instrument-specific marks are located on the rear panel of the instrument next to the nameplate. Operating temperature range: +10 °C to +35 °C Temperature range during storage: +5 °C to +55 °C Relative humidity: max. 80 % non-condensing Storage humidity: < 80 % Section thickness range: 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: approx. 24 mm, ± 2 mm Vertical stroke: 70 mm Max. cutting range without retraction: 69 mm (without specimen orientation 1 µm) Maximum sectioning area with retraction: 62 mm The specimen retraction can be turned off manually: approx. 40 µm

Dimensions and weight Width (including handwheel): Depth (including waste tray): Height (total): Working height (knife blade):

413 mm 618 mm 305 mm (with tray on the hood) 168 mm (measured from the table)

Weight (without accessories):

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 Instructions for Use V 1.8 RevC – 09/2013

4. 4.1

Instrument Setup

Standard delivery

The Leica RM2235 standard delivery includes: 1 Leica RM2235 basic instrument 1 handwheel, complete ����������������������������������������������������������������� 14 0500 38181 1 section waste tray ����������������������������������������������������������������������� 14 0502 37931 1 tool set, consisting of:������������������������������������������������������������������ 14 0500 38600 1 Allen key with handle No. 5�������������������������������������������������� 14 0194 04760 1 Allen key with handle No. 4�������������������������������������������������� 14 0194 04782 1 Allen key No. 3 ����������������������������������������������������������������������� 14 0222 04138 1 screwdriver 3x50, 186 long���������������������������������������������������� 14 0170 11568 1 bottle (50 ml) of oil for drives, type 405�������������������������������� 14 0336 06086 1 brush w/magnet���������������������������������������������������������������������� 14 0183 40426 1 dust protective cover������������������������������������������������������������������� 14 0212 30350 1 Instructions for Use EN... 14 0500 80101 1 language CD... 14 0500 80200 The accessories ordered are included in a separate box. Carefully check the delivery against the packing list and the delivery note. Should you find any discrepancies, please contact your Leica sales office without delay.

4.2

Installation site requirements

• Stable, vibration-free laboratory table with horizontal, flat table top, as far as possible vibration-free ground. • No other instruments nearby which might cause vibrations. • Room temperature consistently between + 10 °C and + 35 °C. • Obstruction-free access to the handwheel. Never operate the instrument in rooms with an explosion hazard.

Leica RM2235

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4.

Instrument Setup

4.3

Unpacking and installation When the instrument is delivered, check the tilt indicators on the packaging. If the arrowhead is blue, the shipment was transported laying flat, was tilted at too great an angle or fell over during transport. Note this on the shipping documents and check the shipment for possible damage. Fig. 6 1

• Loosen and unscrew the six upper screws (2). • Remove the cover (1). • Take the accessory carton (optional accessories) (3) and the cartons (4) from the standard scope of delivery.

2

The transport crate and included retaining elements should be kept in case a return shipment is necessary later.

3

4 5

4 4

4

Fig. 7

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Instructions for Use V 1.8 RevC – 09/2013

4.

Instrument Setup

4.3 Unpacking and installation (cont.) •

• 5

6

Take out the fixing module (5). To do so, hold it by the top edge of the module and in the recessed grip (6) and pull it out by pulling upwards. Lift the instrument* (7) by holding it by the base plate at the front and under the instrument on the back and lift it out of the formed cushion (8). Never hold the instrument for transport by the handwheel or the rotary knob for section thickness adjustment.

7

8

•

(*= the instrument illustration is provided as an example only.)

•

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

•

9 Fig. 8 Leica RM2235

Observe the correct resting angle to the table to avoid pinching your fingers.

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4.

Instrument Setup

4.4

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 in the delivery. 4 3 1

2

2a Fig. 9

16

The feather key (4) is loosely placed in the handwheel shaft (1) and fixed in place with a cable tie 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 a Allen key No. 4 (5). • Remove the cover foil from the self-adhesive cover disk and fix the cover disk on the handwheel.

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Instructions for Use V 1.8 RevC – 09/2013

4. 4.5

Instrument Setup

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

60

12

You can use the quick clamping system (64) to hold all available accessory specimen clamps (for more information, see Chapter 6 "Optional accessories"). To do so, proceed as follows: • Move the specimen head (60) to the upper end position by turning the handwheel (12) and engage the handwheel lock.

60

• To release the clamping system, turn the screw (61) of the quick clamping system (64) counterclockwise using an Allen key No. 4 (71).

62

• 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.

63 64

61

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

71

Fig. 10 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.

Leica RM2235

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4.

Instrument Setup

4.6

Inserting the knife holder

Setting up the knife holder base • 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 sectioning 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. 11

56

71

57

Inserting the knife holder • Loosen the screw (58) using an Allen key No. 4 (71) until 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). • To clamp, retighten the screw (58).

58 Fig. 12

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Instructions for Use V 1.8 RevC – 09/2013

5. 5.1

Operating elements and their functions

5.1.1 Section thickness setting 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

Operation

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. 13

Fig. 14 Leica RM2235

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 specimen - 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.

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