Leica CM1900 Cryostat Instruction Manual V5.3 Oct 2006

1.

Introduction

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 Heidelberger Str. 17 - 19 D-69226 Nussloch

© Leica Microsystems Nussloch GmbH

Tel.: 0 62 24 - 143 0 Fax: 0 62 24 - 143 200 eMail: [email protected] Internet: http://www.histo-solutions.com Leica CM1900 – Cryostat

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2. Table of contents 2. 3. 4. 5.

6.

7.

8.

9.

10. 11. 12. 13. 14. 15. 16. 17. 18.

4

Table of contents ... 4 Safety instructions for handling the instrument ... 5 Technical data ... 8 General description ... 10 5.1 Optional models ... 10 5.2 Designed use/improper handling ... 10 5.3 Standard delivery ... 10 Unpacking the instrument ... 11 6.1 How to open the crate ... 11 6.2 How to remove the packing materia ... 11 6.3 Mounting the ramp ... 12 6.4 Transport to the installation site ... 13 Installation ... 14 7.1 Site requirements ... 14 7.2 Transport to the desired site - Relocation ... 15 7.3 Assembly of the handwheel ... 17 7.4 Locking the handwheel ... 17 7.5 Mounting the heat extractor ... 17 7.6 Inserting the accessories ... 18 7.7 Inserting the optional accessories ... 19 Operation ... 20 8.1 Precooling the knife ... 20 8.2 Installing the knife holder base ... 20 8.3 Inserting the knife holder ... 21 8.4 Connection to the mains ... 21 8.5 Turning on the instrument ... 21 8.6 Leica CM1900 – Overview ... 22 8.7 Programming the desired values ... 24 Daily operation ... 30 9.1 Selection of the adecuate chamber temperature ... 30 9.2 Specimen freezing ... 30 9.3 Activating / deactivating the specimen cooling ... 31 9.4 Activating / deactivating the cryochamber cooling ... 31 9.5 Inserting the specimen discs in the specimen head ... 31 9.6 Inserting the knife in the knife holder ... 32 9.7 Moving the specimen towards or away from the knife via coarse feed ... 33 9.8 Trimming ... 33 9.9 Adjustment of the anti-roll guide ... 34 9.10 Sectioning ... 34 Cleaning and disinfection ... 35 10.1 Cleaning and disinfection ... 35 10.2 Turning the instrument back on ... 35 Removal of the microtome ... 36 11.1 How to remove the microtome: ... 36 Reinstallation of the microtome ... 38 12.1 How to return the microtome to the cryochamber ... 38 Maintenance ... 40 13.1 General maintenance ... 40 13.2 Replacement of the lamp ... 41 Troubleshooting ... 42 Temperature Selection Chart ... 45 Optional accessories ... 46 16.1 Orienting specimen head ... 46 16.2 Thermal block ... 47 Ordering information ... 48 Warranty and service ... 50

Instruction manual V 5.3 – 10/2006

3.

Safety instructions for handling the instrument

Unpacking and installation

Microtome knives

• To ensure an adequate cooling capacity, the instrument must be set up maintaining a minimum distance from walls and furniture (see 'Technical data'). • The instrument must be transported in an upright position or slightly tilted (max. 30°). • To ensure a safe transportation with a fork lift 3 people are required: one operating the fork lift, and the other 2 holding the instrument on either side to prevent it from sliding down. • Before connecting to the mains power, please check if the local voltage complies with the power rating specified on the name plate of the instrument (see also 'Technical data')! • Plug the instrument only to power sockets with ground! • After transporting, wait at least 4 hours before turning the instrument on. This waiting period is necesssary to allow the compressor oil , which may have been displaced during transport, to return into its original position. Failure to comply with this will cause severe damage to the instruments!

• Take care when handling microtome knives and disposable blades. The cutting edge is extremely sharp and can cause severe injury! • Never leave knives and knife holders with a knife/ blade mounted lying around! • Do not place a knife on a table with the cutting edge facing upward! • Never try to catch a falling knife! • Prior to manipulating the knife and specimen, or changing the specimen or knife, always lock the handwheel and cover the cutting edge with the knife guard! • Always lock the handwheel and cover the cutting edge of the knife with the knife guard during breaks! • Avoid contact with cold parts of the instrument as this can cause frostbite!

Due to the weight of the instrument (170 kg) 4 people are required for transportation (2 people per handle).

Symbols used in this manual and their meaning Warnings appear in a gray box and are marked by a warning triangle

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

(5) (Fig.5)

Figures in brackets refer to item numbers in drawings or to the drawings themselves.

Leica CM1900 – Cryostat

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3.

Safety instructions for handling the instrument

Cleaning and disinfection - Turning the instrument back on It is not necessary to remove the microtome for disinfecting the cryochamber, however, removal is possible, if required!

• When disinfecting, please take appropriate protective measures (gloves, mask, protective clothing, etc.). • When using detergents and disinfectants please comply with the safety precautions of the disinfectant manufacturer. • Only use acetone for cleaning the plastic anti-roll plate of the knife holders CN and CS. The glass antiroll plate of the knife holder CE can be cleaned either with acetone or alcohol. • Dispose of waste liquid according to the waste disposal regulations! • Do not use uncontrolled external heaters for drying the cryochamber. This can cause damage to the cooling system!

Removal/Reinstallation of the microtome • Prior to removing the microtome, turn the instrument off with the mains switch and pull the mains plug! • Remove sliding window before removing the microtome! • Before removing the microtome, lock the handwheel in the lowest position. When removing the microtome, the specimen head will rapidly fall down and might injure the operator's hands! • The microtome must be entirely dry before reinstallation. Humidity inside will condense and freeze, causing malfunctions and damage! • All components removed from the cryostat must be carefully dried before returning them to the cryochamber! • The cooling chamber must be entirely dry when turning on the instrument (Frost formation)! • When removing the microtome for cleaning or disinfection, please keep in mind the safety instructions in chapter 10 'Cleaning and Disinfection'.

• Do not turn the instrument on before the cryochamber is completely dry! Frost formation! • Dry all parts completely before reinserting them in the cryochamber! • The front panel and the slit cover of the microtome must be completely dry before turning on the instrument!

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Instruction manual V 5.3 – 10/2006

3.

Safety instructions for handling the instrument

Handling samples - Defrosting

Maintenance

• When working with possibly contaminated or infected material the general safety guidelines for laboratories must be applied!

• Turn the instrument off with the mains switch and pull the mains plug, before replacing the fuses!

• Before defrosting the cryochamber remove all samples! • Before defrosting the specimen head remove all samples! • Never leave samples in the cryochamber! - The instrument is not made for storing frozen specimen!

• Only use fuses of the same specification ! For the required values refer to chapter 4 'Technical Data'! • Turn the instrument off with the mains switch and pull the mains plug before replacing the lamp! • If the lamp is broken, it must be replaced by the technical service, as the replacement involves a high risk of injury! • Only use lamps of the same specification! For the required type refer to chapter 4 'Technical Data'!

Leica CM1900 – Cryostat

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4. Technical data Type

-1

-2

-3

-4

-5

-6

Nominal voltage Nominal frequency Power draw Max. start-up current for 5 sec Protective class Mains fuses (MDA by Bussmann) Automatic fuse Pollution degree ➁ Overvoltage installation category Heat emission (max.)

230 V AC 50 Hz 1800 VA 25 A eff. I T8A T10A T1 2 II 1800 J/s

120 V AC 60 Hz 1800 VA 35 A eff. I T15A T1 2 II 1800 J/s

230 V AC 60 Hz 1800 VA 25 A eff. I T8A T10A T1 2 II 1800 J/s

240 V AC 50 Hz 1800 VA 25 A eff. I T8A T10A T1 2 II 1800 J/s

100 V AC 60 Hz 1800 VA 30 A eff. I T15A M3 2 II 1800 J/s

100 V AC 50 Hz 1800 VA 35 A eff. I T15A M3 2 II 1800 J/s

Refrigeration Cryochamber Temperature range

CM1900, 50 Hz

CM1900, 60 Hz

0 - -35 °C ± 3 K, at an ambient temperature of 22 °C

0 - -35 °C ± 3 K, at an ambient temperature of 22 °C

Refrigeration capacity ➀ Cut-off pressure Safety factor Refrigerant* Compressor oil*

690 W 25 bar 3 275 g (± 5 g) refrigerant R 404A* 0,6 l EMKARATE RL22S, ICI*

690 W 25 bar 3 265 g (± 5 g) refrigerant R 404A* 0,6 l EMKARATE RL22S, ICI*

Defrosting of cryochamber Automatic defrosting Programmable: Defrosting intervals Defrosting period: Automatic stop of manual defrosting:

yes 1 defrost cycle/ 24 hours 9 minutes at -5 °C

yes 1 defrost cycle/ 24 hours 9 minutes at -5 °C

Manual defrosting Defrosting period: Automatic stop of manual defrosting:

9 minutes at -5 °C

9 minutes at -5 °C

Quick freeze shelf Max. temperature: Number of quick freeze stations:

- 43 °C (+ 0 K / - 2 K) 10

- 43 °C (+ 0 K / - 2 K) 10

-10 - -50 °C ± 2 K, at an ambient temperature of 22 °C

-10 - -50 °C ± 2 K, at an ambient temperature of 22 °C

Refrigeration capacity ➀ Cut-off pressure Safety factor

320 W 25 bar 3

320 W 25 bar 3

Refrigerant* Compressor oil*

210 g (± 5 g) refrigerant R404A* 0,4 l alpha 22, Kyodo*

210 g (± 5 g) refrigerant R404A* 0,4 l alpha 22, Kyodo*

Defrosting of specimen head Automatic defrosting

no

no

Manual defrosting Defrosting period: Automatic stop of manual defrosting:

10 minutes 10 min. after reaching +20 °C

10 minutes 10 min. after reaching +20 °C

Specimen cooling Temperature range

➀ acc. to CECOMAF: liquid temperature 45 °C, evaporation temperature: -25°C

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➁ according to IEC-1010; UL 3101

Instruction manual V 5.3 – 10/2006

4.

Technical data

Microtome Type Section thickness setting Specimen feed Knife holder repositioning on the microtome base plate Vertical stroke Specimen retraction Maximum specimen size

Microtome encapsulated in the cryochamber 0 - 60 μm, infinitely variable 25 mm 30 mm 59 mm yes (US optional) Ø 40 mm

Cryostat Dimensions Width (w/o handwheel) Width (incl. handwheel) Depth (cabinet only) Depth (cabinet incl. mains plug) Overall height Working height

790 mm 890 mm 725 mm 800 mm 1200 mm 1000 mm

Weight (instrument only) Weight incl. accessories

168 kg 170 kg

Set-up conditions Distance to walls and furniture; calculated from the cabinet

rear: right side: left side:

10 cm 20 cm 10 cm

Plug the instrument only to power sockets with ground. Mains cable length: – 3,5 m Extendable: no The installation site must be free of draft and insolation; the floor must be free from vibrations.

Lamp 50 Hz-Version: 60 Hz-Version:

Conditions ambient temperature: 8 °C to 40 °C Operating temperature range: 0°C to -35 °C Temperature range during storage: + 5°C to +55°C Relative humidity: max. 60%, non-condensing Humidity during storage: < 60%

Leica CM1900 – Cryostat

Osram DULUX S 11 W/21 Osram DULUX S 13 W/21

*) Refrigerant and compressor oil must only be replaced by authorized service personnel!

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5.

General description 5.1

Designed use/improper handling

The rapid sectioning cryostat CM1900 is provided with a fully encapsulated microtome with an independent specimen cooling system. The cryostat CM1900 is appropriate for InVitro Diagnostic (IVD) applications. The instrument may only be operated within the scope of its designated use as described above and as per the instructions given in this manual. The instrument is CFC-free and is designed and manufactured according to VDE and UL regulations.

5.2

Optional models

The following models are available: • CM1900 with specimen retraction • CM1900 without specimen retraction Please, indicate type when ordering!

5.3

Samples must not be stored unattended in the cryostat for a long period since they could be destroyed in case of a power failure or malfunction of the instrument. The cryochamber temperature can rise considerably during defrosting. Heat-sensitive samples must therefore be protected from exposure to elevated temperatures and removed prior to defrosting. The instrument may only be used for the specified application and operated in accordance with the instructions given in this manual. Any other use of this instrument is considered as improper operation!

Standard delivery 1 Basic instrument 1 Handwheel, assy. ... 0416 18478 1 Heat extractor, stationary ... 0452 27918 1 Low-temperature stabilizer for heat extractor ... 0452 27919 1 Set of specimen discs ... 0470 43550 - 4 specimen discs, 25 mm ... 0416 19275 - 4 specimen discs, 30 mm ... 0370 08587 1 One-piece storage shelf ... 0452 28594 1 Freezing shelf cover ... 0452 28624 1 Brush shelf with holder ... 0452 28799 1 Section waste tray ... 0452 28595 1 toolkit: ... 0436 43463 - 1brush, fine ... 0183 28642 - 1 Leica-brush ... 0183 30751 - 1 Allen key, no. 1.5 ... 0222 10050 - 1 Allen key, no. 2.5 ... 0222 04137 - 1 Allen key, no. 3.0 ... 0222 04138 - 1 Allen key, no. 4.0 ... 0222 04139 - 1 Allen key with spherical head, no. 4.0 ... 0222 32131 - 1 Allen key, no. 5 ... 0222 04140 - 1 Allen key with handle, no. 5 ... 0194 04760 - 1 Allen key, no. 6 ... 0022 04141 - 1 Single-head wrench, no. 13 ... 0330 33149 - 1 Single-head wrench, no. 16 ... 0330 18595 1 Bottle of OCT-Compound, mounting med. f. cryosectioning, 125 ml ... 0201 08926 1 Bottle of cryostat oil, type 407, 50 ml ... 0336 06089 1 Storage mat ... 0452 27913 1 Pair of safety gloves for cryosectioning, size S ... 0340 40859 1 Instruction manual Leica CM1900 - G/E/F/S ... 0708 37105

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Instruction manual V 5.3 – 10/2006

6.

Unpacking the instrument

Checking if the instrument has been transported correctly

Please check the TIP (N) TELL indicators immediately after arrival of the shipment. If the TIP (N) TELL Arrow Point is blue, this package has been on its side or tipped in transit. Please note on the Bill of Lading and check for damage. The same unpacking instructions are inside a transparent sheet, attached to the transport crate at arrival of the shipment. The following unpacking instructions shall help you in case of packing or transporting the instrument once again.6.1

6.1 1 1

How to open the crate

1. Loosen 3 screws (1) at the top of the two sides of the crate (7).

1 3

2

2. Loosen 4 screws of the hinges (2) at the front and rear of the crate.

2 7

Do not loosen the screws of the hinges on the lid of the crate!

4

3. Loosen 1 screw (3) at the top between the two hinges at the front and rear of the crate.

4 4

4

4

4

5

4. Loosen 3 screws (4) at the bottom on each of the four sides of the crate.

6.2

1. Remove the lid (5) and place on the ground beside the transport platform (11) with the foam parts facing down.

6

6

How to remove the packing materia

2. Remove the 4 foam parts (6).

7

3. Lift the body of the crate (7) straight up.

9

4. Take out the cardboard box (8) containing the accessories and the instruction manual at the front of the instrument. 10

8

5. Remove the dust cover (9) by pulling it upward. 6. Remove the foam part (10) at the rear.

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Leica CM1900 – Cryostat

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6.

Unpacking the instrument 6.3

11

Mounting the ramp

1. Upon removal of the foam part (10) 2 screws (12) become visible on the base platform.

12

2. Fold down the hinges (2) at the rear and front. 3. Place the lid against the transport platform (11) from behind. The jogs (13) of the hinges (2) must point to the instrument. 2

13

4. Push the lid to the left sliding the jogs (13) of the hinges under the heads of the screws (12).

11

12 13

12

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Instruction manual V 5.3 – 10/2006

6. 6.4

Unpacking the instrument

Transport to the installation site The instrument must be transported in an upright position or slightly tilted (max. 30°)!

16

• The lid (5) forms a ramp for the instrument so it can be rolled off the transport platform. Do not grip the cabinet at the lid (16)! Grip the cabinet only at the marked locations ( )! 2

2

14

The wheels (14) must be guided over the hinges (2) at the front and rear. Caution: Risk of tilting!

1. Roll the instrument backwards over the ramp off the platform with caution.

5

2. Move the instrument to the installation site on its wheels. 3. The adjustable feet (15) can support the weight of the instrument when tipped at a slight angle (max. 30°). Tilt

Lengthwise movement

15 14 15

Leica CM1900 – Cryostat

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7.

Installation

7.1

Site requirements To ensure an adequate cooling capacity, the instrument must be set up maintaining a minimum distance from walls and furniture (see 'Technical data')!

The place of installation must meet the following requirements: • No direct sunlight. • Mains power socket at a distance no greater than appr. 3 m. • No drafts (air condition outlets etc.). • Even floor. • Vibration-free floor. • Only for indoore use. • Obstruction-free access to the handwheel. • Obstruction-free access to the ON/OFF switch. • Room temperature always approx. 22 °C. • Air humidity must not exceed 60 %. • Distance to walls and furniture, calculated from the cabinet: - rear: 10 cm - right side: 20 cm - left side: 10 cm • No heat dissipating appliances around. High room temperatures and excessive air humidity affect the cooling capacity of the cryostat and lead to the formation of condensation water in the instrument!

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Instruction manual V 5.3 – 10/2006

7. 7.2

Transport to the desired site - Relocation

• First, check if the location meets the conditions specified in 'Site requirements'. • Transport the instrument to the desired location. • Observe the following: The instrument must be transported in an upright position or slightly tilted (max. 30°)!

Installation

Transport on wheels • On even floors and for short distances the instrument can be transported on wheels. Before transportation, screw the adjustable feet (1) completely down with the open-end wrench no. 16. • The instrument can be rolled lengthwise and slightly be tilted forward (see arrows). - Although the instrument cannot be rolled sideways, it can be pushed carefully in that direction. When tilting the instrument 2 people must counterbalance from the frontside to prevent the instrument from falling down and causing severe injury!

Tilt

Lengthwise movement

3

2 1

• When transporting the instrument on wheels (2, 3) grip the cabinet only at the marked locations ( ). • The adjustable feet (15) can support the weight of the instrument when tipped at a slight angle (max. 30°). Unscrew the adjustable feet with the open-end wrench no. 16 (when subsequently transporting the instrument any further on wheels, do not forget to screw the adjustable feet down completely).

Leica CM1900 – Cryostat

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7.

Installation

Transport with handles

Transport with a fork lift

• For lifting or carrying the instrument manually (e.g. on stairs) you can use handles (4). Screw the handles into the threads (5) of the bottom plate at both sides of the cabinet. • The handles can be ordered from your Leica Sales Unit.

• The instrument can be transported with a fork lift. • The distance 'a' for insertion between the wheels is max. 540 mm.

4

4

5

Distance for insertion a = max. 540 mm

5

Due to the weight of the instrument (170 kg) 4 people are required for transportation (2 persons per handle).

To ensure a safe transportation with a fork lift 3 people are required: one operating the fork lift, and the other 2 holding the instrument on either side to prevent it from sliding down.

• Unscrew the adjustable feet at the installation site (1) with the open-end wrench no. 16 (see Fig. page 16). This is absolutely necessary for a stable stand.

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Instruction manual V 5.3 – 10/2006

7. 7.3

Installation

Assembly of the handwheel

1 2 3 4

• Insert the pin (1) of the handwheel shaft into the hole (2) of the handwheel. • Mount the spring washer (3) on the screw (4). • Tighten the screw (4) with an Allen key no. 6. To dismount, proceed in reverse order.

7.4

• To unlock, push the locking pin (5) to the left from the recess at the handwheel.

7.5

Mounting the heat extractor

Locking the handwheel 7

6 5

• For locking the handwheel rotate the handle until it is in the upper position. Push the locking pin (5) into the recess at the handwheel. The locking position is marked by a black dot (6). If necessary, move the handwheel slightly forth and back until the locking mechanism engages.

Leica CM1900 – Cryostat

• Fix the holding device of the heat extractor by tightening the 2 screws (7) with the Allen key no. 4 to the threaded holes on the left sidewall of the cryochamber and insert the heat extractor.

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7.

Installation

7.6

Inserting the accessories

• Insert the storing shelf (1). 1

18

• Insert the rimmed section waste tray (2). 2

Instruction manual V 5.3 – 10/2006

7. 7.7

Installation

Inserting the optional accessories

• Insert the waste container (3).

• Insert the right storage shelf (4). 4

3

• Insert the left storage shelf (5).

• Insert the section waste tray (6).

5 6

Leica CM1900 – Cryostat

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8.

Operation

8.1

Precooling the knife

• To precool the knife put the open knife rest including the knife on the storage shelf inside the cryochamber.

8.2

Installing the knife holder base

• To unlock relocate the lever to the front (1).

1

• Slide the guide (2) of the knife holder base (3) onto the T-piece (4) of the microtome base plate as shown.

The knives are extremely sharp! Handle with care! Never try to catch a falling knife!

3 2

4

• To clamp relocate the lever (1) backwards.

1

• If the lever cannot be relocated entirely for clamping, lift it up and shift it to the next position.

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Instruction manual V 5.3 – 10/2006

8. 8.3

Inserting the knife holder

• To insert the knife holder, unlock the lever (1) turning it upward, slightly to the rear. • Insert the knife holder (2) onto the knife holder base (3) as shown.

Operation

• Do not connect other consumers to this electric circuit. • Connect the mains plug to the mains power outlet at the wall. After transporting, wait at least 4 hours before turning the instrument on. This waiting period is necessary to allow the compressor oil, which may have been displaced during transport, to return to its original position. Failure to comply with this can cause severe damage to the instrument!

1

2

8.5

Turning on the instrument

• ON/OFF switch must be easily available. • The automatic mains fuse is used as mains switch. • Bring the switch of the automatic fuse (2) to the upper position. • Turn on the mains switch (1). 3

• Close the sliding window.

• To clamp, turn the lever (1) forward.

8.4

Connection to the mains

During the start-up of the compressor the nominal voltage must not drop below the values specified in the 'Technical Data'. Please note that the compressor requires a start-up current between 30 and 40 A. Therefore, the electric circuit at the installation site must be inspected by an electrical engineer to ensure that it meets the requirements for a troublefree operation of the instrument. Failure to comply with the above will cause severe damage to the instrument!

1

2

To avoid frost formation always put the cover on the quick freeze shelf. Always cover the quick freeze shelf during breaks and overnight.

• Check mains voltage and mains frequency to comply with the specification on the type plate. Plug the instrument only to power sockets with ground!

Leica CM1900 – Cryostat

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