Leica CM1950 Bedienungsanleitung

Operating Manual Leica CM1950 , V 1.1 English – 05/2007 Always keep this manual near the instrument. Read carefully prior to working with the instrument! Leica CM1950 Cryostat[...]

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3 Leica CM1950 – Cryostat Published by: Leica Biosystems Nussloch GmbH Heidelberger Str. 17 - 19 D-69226 Nussloch Germany Phone: +49 (62 24) 143-0 Fax: +49 (62 24) 143-2 00 eMail: histo_info@leica-microsystems.com Internet: http://www.histo-solutions.com These are determined only by the contract provi- sions agreed between ourselves and our custo[...]

4 Operating Manual V 1.1 – 05/2007 Table of contents 1. Important Information ......................................................................................................................... 6 1.1 Symbols Used in This Manual and Their Meaning ................................................................ 6 1.2 Qualification of Personne[...]

5 Leica CM1950 – Cryostat 7. Instrument Controls ........................................................................................................................... 30 7.1 Control Panel Fields on the CM195 0 – Control Panel Field 1 .............................................. 30 Control Panel Field 2 – Electric Coarse Feed, Sectioni[...]

6 Operating Manual V 1.1 – 05/2007 1. Important Information 1.1 Symbols Used in This Manual and Their Meaning Notes i.e. important user information appear in a gray box and are marked by an in- formation symbol. Caution – UVC radiation! Figures in brackets refer to item num- bers in drawings or to the drawings themselves. (5) (Fig.5) Warnings a[...]

7 Leica CM1950 – Cryostat 2.1 General Safety Instructions The CM1950 is a cryostat with an encapsulated microtome and separate specimen cooling. It is primarily used for work in the area of fast-cut diag- nostics. The displays and instrument controls are easy to operate due to their largely self-explanatory symbols. LED displays make it easy to r[...]

8 Operating Manual V 1.1 – 05/2007 2. Safety and Design Opening the packaging 1. Remove the 3 screws ( 1 ) on each of the two sides of the box ( 7 ) at the top. 2. Remove the 2 screws on each of the 4 hinges ( 2 ) on the front and rear side of the box. Do NOT remove the screws of the hinges on the top lid! 3. Remove 1 screw ( 3 ) between the two [...]

9 Leica CM1950 Cryostat 2. Safety and Design Removing the packaging 1. Remove the cover ( 17 ) and place it on the floor with the foam parts facing downwards. - The cover will be used as a ramp later. 2. Remove the 4 foam parts ( 6 ) by pulling them upwards. 3. Open the box ( 7 ) on its side hinges like an accordion partition and remove it. 4. Remo[...]

10 Operating Manual V 1.1 – 05/2007 To prevent adverse health effects from UV radiation, the UV disinfec- tion cycle can be started only after the sliding window has been prop- erly closed. Closing the window activates the corresponding safety features. The consistent use of these safety features and strict observation of the warn- ings and cauti[...]

11 Leica CM1950 – Cryostat 2.4 Securing/Locking the Handwheel Always lock the handwheel and cover the cutting edge with the knife guard prior to manipulating the knife or specimen, changing the specimen, or taking a break. To lock the handwheel, press the lever ( 1 ) outward. Continue turning the handwheel slowly until the grip is in the upper or[...]

12 Operating Manual V 1.1 – 05/2007 2. Safety and Design • When disinfecting the instrument, take appropriate protective measures (gloves, mask, protective clothing, etc.). • When using detergents and disinfectants please comply with the safety precautions of the disinfectant manufacturer. • The integrated glass anti-roll guide of the blade[...]

13 Leica CM1950 Cryostat 2. Safety and Design 2.7 Removal of the Microtome • The microtome is encapsulated and therefore does not require removal by the user. 2.8 Maintenance Replacing the fuses • Turn the instrument off and disconnect the power plug before replacing the fuses. • Use only the fuse types specified in chapter 3, "Technical[...]

14 Operating Manual V 1.1 – 05/2007 3. Technical Data ② according to IEC-1010; UL 3101 Type Nominal voltage (±10 %) Nominal frequency Power draw Max. start-up current for 5 sec. Protective class Circuit breaker: (MDA made by Schurter) Pollution degree ② Overvoltage installation category Heat emission (max.) Approval -1 -2 -3 100 V AC 120 V A[...]

15 Leica CM1950 – Cryostat 3. Technical Data Refrigeration system CM1950 , 50 Hz CM1950 , 60 Hz Cryochamber Temperature range 0 °C to -35°C ± 5 K, adjustable in 1K increments, 0 °C to -35°C ± 5 K, adjustable in 1K increments at ambient temperature of 20 °C ambient temperature of 20 °C Cooling time to -2 5 °C approx. 5 h approx. 5 h Cooli[...]

16 Operating Manual V 1.1 – 05/2007 4. Standard Delivery A choice of different knife holders is available for the CM1950 . Compare the delivered components with the parts list and your order. Should you find any discrepancies, please contact your Leica sales office without delay. Basic instrument WITHOUT motor/WITHOUT extraction, in the specific [...]

17 Leica CM1950 – Cryostat 4. Standard Delivery Basic instrument WITH motor/WITHOUT extraction, in the specific voltage variant 1 handwheel, motorized .................................................................................... 0477 41347 5 specimen discs, 30 mm .............................................................................[...]

18 Operating Manual V 1.1 – 05/2007 5. General Overview Castors for safe transport over short distances Handwheel in 12 o’clock position On/off switch, also circuit breaker Align adjusting feet for secure upright position Unscrew and align adjusting feet after transport Bacterial filter (optional, only for instruments with filter) Catch tank fo[...]

19 Leica CM1950 – Cryostat 5. General Overview 5.1 Control Panel Fields and Cryostat Chamber 1 2 3 1 - Control panel field 1: Extraction, temperature and time control, illumination, UV disinfection 2 - Control panel field 2: Electric coarse feed (sectioning and trimming thickness adjustment) 3 - Control panel field 3: Motorized sectioning cutting[...]

20 Operating Manual V 1.1 – 05/2007 6. Installation 6.1 Site Requirements The place of installation must meet the following requirements: • No direct sunlight. • Power supply at a distance no greater than approx. 3 m. • No drafts (air condition outlets etc.). • Even floor. • Vibration-free floor. • The instrument is designed for indoo[...]

21 Leica CM1950 – Cryostat 6. Installation 6.2 Transport to the Site • 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 slight- ly tilted (max. 30°)! • When[...]

22 Operating Manual V 1.1 – 05/2007 • Insert the pin ( 1 ) of the handwheel shaft into the hole ( 2 ) of the handwheel. • Tighten the screw ( 4 ) using the No. 6 Allen key. • Place the protective cap on the screw ( 4 ). To dismount, proceed in reverse order. 1 2 4 6.3 Assembling the Handwheel Fig. 14 Transport with a fork lift • The instr[...]

23 Leica CM1950 Cryostat 6.3.1 Locking/Unlocking the Handwheel Only rotate the handwheel when the refrigeration system is on and the cryochamber is cold! Always lock the handwheel prior to manipulating the knife or specimen, changing the specimen, or taking a break. To lock the handwheel, move its handle to the 12 or 6 o’clock position. Press the[...]

24 Operating Manual V 1.1 – 05/2007 6.4 Connection to Power Supply System 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 re- turn into its original position. Furthermore, any condensation that has formed du[...]

25 Leica CM1950 – Cryostat 6.5 Installing Accessories/Inserting Chamber Accessories 6.5.1 Installing the Adjustable Footrest (Optional) • To install the optional footrest, unscrew the screws ( 1 ) using the No. 3 Allen key provided. 1 • Attach the holders ( 2 ) for the footrest on the left and right to the front wall of the housing. To do so,[...]

26 Operating Manual V 1.1 – 05/2007 6.5.2 Installing the Storage Systems (Optional) 6.5.3 Shelf, Movable (Optional) Attach the rod for the shelf to the inner front side of the cryostat housing using the provided screws ( 1 ) and the No. 3 Allen key, then attach the caps ( 3 ). (The rear side of the movable shelf has white plastic screws ( 2 ) tha[...]

27 Leica CM1950 – Cryostat 6. Installation 6.5.5 Installing the Heat Conductor Block, Stationary (Optional) Fig. 24 6.5.6 Installing the Knife/Blade Holder and Adjusting the Clearance Angle For temperature reasons, install the knife/blade holder on an appropriate base. • Set the knife or blade holder ( 1, Fig. 25) on the base ( 2 ), adjust the [...]

28 Operating Manual V 1.1 – 05/2007 Reasons: 1. To prevent section waste from falling into the opening. 2 . To prevent cold from escaping from the chamber. 3. To prevent moisture from penetrating into the chamber. 6. Installation 6.5.7 Inserting/Changing the Bacteria Filter 6.5.8 Assembling the Filter Bag Fig. 27 • Set the mark ( A ) of the ext[...]

29 Leica CM1950 – Cryostat 6.5.9 Installing the Section Extraction (Optional) – Use with Blade Holder CE Only 6. Installation If the extraction is not used for a long time, it is absolutely necessary to clean the extraction hose in order to ensure maximum extraction capacity. To do so, place the hose in commercially available disinfectant or al[...]

30 Operating Manual V 1.1 – 05/2007 7. Instrument Controls Display of actual and target specimen head temperatures Display of actual and target chamber temperatures Display of real time, defrost time and error messages Max-Cool button to select the maximum low temperature of the specimen head directly (-50 °C) +/- buttons to select specimen head[...]

31 Leica CM1950 Cryostat • The Peltier element provides additional cooling for the freezing stations. After the button is pressed, the display changes from "PE" to 10", indicating an additional cooling period of 10 minutes. The remaining cool- ing time is always shown in the display. Once only 4 minutes are remain- ing, a dot will [...]

32 Operating Manual V 1.1 – 05/2007 For trimming values above a sec- tion thickness of 200 μm, the dis- play flashes to expressly notify the user of thick sections! Control Panel Field 2 – Electric Coarse Feed, Sectioning and Trimming Thickness Display 1 Specimen head forwards – pressing briefly advances the specimen 20 μm Specimen head fas[...]

33 Leica CM1950 – Cryostat 7. Instrument Controls Coarse feed functions The two-speed electric coarse feed is designed to quickly position the spec- imen in relation to the knife. The buttons with double arrows move the spec- imen at 900 μm/s; the buttons with single arrows move it at 300 μm/s. Retracting the specimen head from the knife • Pr[...]

34 Operating Manual V 1.1 – 05/2007 Sectioning modes The instrument can be used in manual and motorized modes. The following settings are available: • Single stroke (SINGLE) or continuous stroke (CONT .) in automatic mode, as well as • ROCK (sectioning using handwheel) in manual mode. When switching the instrument on, no operating mode is act[...]

35 Leica CM1950 – Cryostat 7. Instrument Controls Disinfection Duration - 30 min Duration - 180 min UVC key - to activate / deac- tivate the disinfection cycle and/or to acknowledge in- terruption of a disinfection cycle. To start disinfection the sliding window must be completely closed. • Press UVC key once briefly to start the 30 min. mode ?[...]

36 Operating Manual V 1.1 – 05/2007 8. Working with the Instrument 8.1 Preparing Cutting Tools, Specimen Discs and Preparation Aids The knives are extremely sharp! Handle with care! Never try to catch a falling knife! • Place working materials such as the blade box or knives (in the knife case), brush, pincers or preparation needles and, where [...]

37 Leica CM1950 Cryostat 8. Working with the Instrument 8.3 Configuring the Parameters The instrument must be switched on at least 5 hours before the planned use. • Turn the lamp on or off. • This button activates or deactivates the manual defrost of the cooling chamber, specimen head or freeze shelf. (For more detailed handling instructions, r[...]

38 Operating Manual V 1.1 – 05/2007 Setting the time • The clock time is set with the +/- buttons in the control panel marked with the clock symbol. To do so, set the current time using the "+" or "-" button below the small clock symbol. Pushing the "+" or "-" button for more than 1 sec. increases or decr[...]

39 Leica CM1950 Cryostat 8. Working with the Instrument Programming the defrost cycle • Set the beginning of the automatic defrost cycle. The automatic defrost cycle takes place once within 24 hours. It is set with the "+"/"-" buttons on the right of the panel with the clock symbol. The two buttons are marked by a melting snow[...]

40 Operating Manual V 1.1 – 05/2007 8. Working with the Instrument Manual defrosting of the cryochamber • Push the manual defrost button (with the melting snowflake) on the left over the key button to activate the defrost cycle of the cryochamber on demand. • Activation is confirmed by an audible signal. • Then, push the "+" or &q[...]

41 Leica CM1950 Cryostat 8. Working with the Instrument Entering the trimming thickness To switch from a trimming section thickness for research applica- tions (1-600 μm) to a thickness for clinical applications (10, 20, 30 or 40 μm), press and hold down the TRIM/SECT key while switching on the instrument. • Press the TRIM/SECT button. TRIM mod[...]

42 Operating Manual V 1.1 – 05/2007 8. Working with the Instrument 8.4 Working with the Precooled Cryostat 8.4.1 Preparatory Work • Lock the handwheel in the top (12 o’clock) position. • Cut the specimen to size outside of the cryostat. • Select the precooled specimen disc, cover it with freezing compound, then attach and orient the speci[...]

43 Leica CM1950 – Cryostat 8. Working with the Instrument 8.4.2 Trimming with Extraction – Anti-roll Guide Installed • Remove the silicone stopper from the filter cover (and keep it in a safe place). • Insert the extraction hose with the black adapter. • Fold the anti-roll guide to the side and fasten the suction nozzle to the pressure pl[...]

44 Operating Manual V 1.1 – 05/2007 8. Working with the Instrument For motorized sectioning, always begin at low speed for safety reasons. • Optimize the vacuum settings if necessary. • Press the "VAC" button to enable the vacuum extractor. The LED in the "VAC" button is lit while the extractor is on. Press the button agai[...]

45 Leica CM1950 Cryostat 8. Working with the Instrument • Activate trimming mode. • Select the trimming thickness. • Switch on the VAC and select a low extraction level (between 1 and 2). • Start trimming by manually moving the handwheel or press the RUN/STOP and ENABLE buttons simultaneously to start motorized sectioning. • Use a brush t[...]

46 Operating Manual V 1.1 – 05/2007 A few rules: • Always begin at a low extraction level, then slowly increase it. • Do not use high extraction levels unless absolutely necessary. • Different specimen sizes require different extraction levels. • The faster the trimming or sectioning speed, the lower the extraction level should be. • Th[...]

47 Leica CM1950 Cryostat 8. Working with the Instrument After trimming or sectioning: Specimen: • Unclamp and thaw. • Immerse in fixative for further processing. Cleaning • Use the brush to sweep up the section waste (section waste tray) and remove it from the cryostat (follow applicable laboratory regulations for disposal). Or: • Clean the[...]

48 Operating Manual V1.1 – 05/2007 Problem Causes Remedies Frost on chamber walls and microtome Sections smear Sections splinter Sections not properly flattened Sections not properly flattened despite correct temperature and correctly aligned anti-roll plate Sections curl on anti-roll plate Scraping noise during sectioning and specimen return mov[...]

Leica CM1950 Cryostat 49 Problem Causes Remedies Ridged sections Chatter during sectioning Condensation on anti-roll plate and knife during cleaning Anti-roll plate damaged after adjustment Thick-thin sections Knife damaged. Edge of anti-roll plate damaged. Specimen insufficiently frozen onto the specimen disc. Specimen disc not clamped tightly. Kn[...]

50 Operating Manual V1.1 – 05/2007 Problem Causes Remedies Blunt knife. Inappropriate section thickness. Incorrect clearance angle. Microtome not dried thoroughly enough. Dried specimen. Anti-roll plate is too warm or incorrectly positioned. Fat on the corner or edge of anti-roll plate. Anti-roll plate not correctly fixed. Rust on the knife. Anti[...]

Leica CM1950 Cryostat 51 Problem Causes Remedies No or insufficient refrigeration of the cryochamber Sliding window collects condensation No or insufficient refrigeration of the specimen Lamp does not work Both disinfection LEDs flash alternately Cooling system or electronic drive defective. Air humidity and room temperature too high. Cooling syste[...]

52 Operating Manual V 1.1 – 05/2007 10. Temperature Selection Chart (in minus °C) Tissue type Chamber temperature Specimen head temperature Spleen -15 °C to -20 °C -11 °C Liver -10 ° C -20 ° C -15 °C off to -15 °C Intestine -10°C -20 °C -15 °C A*: off to -20 °C E*: -20 °C Heart -10 °C A: -20 °C E: -20 °C to -30 °C -15 °C off to [...]

53 Leica CM1950 – Cryostat 11.1 Ordering Information Specimen disc ø 20 mm assembly .................................................... 0477 43739 Specimen disc ø 30 mm assembly .................................................... 0477 40044 Specimen disc ø 40 mm assembly .................................................... 0477 40045 Specime[...]

54 Operating Manual V 1.1 – 05/2007 Section waste tray .............................................................................. 0477 40062 Brush shelf ........................................................................................... 0477 43036 Storage system, assembly ...............................................................[...]

55 Leica CM1950 – Cryostat Assembling the anti-roll guide system (for the blade holder CE) 1. Insert the glare-minimizing glass insert into the interchangeable frame and tighten it evenly using the knurled screw ( 10 ). 2. Insert the shaft ( 11 ) of the metal frame for exchangeable glass inserts from above into the bore of the swinging arm ( 12 )[...]

56 Operating Manual V 1.1 – 05/2007 The knife guard is folded away in the detail view. Knife guard folded away Fig. 43 Four possible settings Blade rest (ruler) for low-profile blades 1 2 1 2 6 Fig. 44 11. Optional Accessories Red elements on the blade or knife holders, such as the knife guard and ejector, are protective devices that may not be r[...]

57 Leica CM1950 – Cryostat Anti-roll guide system functions as a knife guard here! 1. Fold the anti-roll guide system ( 4 ) to the left – while doing so, hold the lever ( 11 ) ( not the adjusting screw of the anti-roll guide) so that the height of the anti-roll guide remains unchanged. 2. Open the clamping lever ( 10 ) by turning it coun- tercl[...]

58 Operating Manual V 1.1 – 05/2007 11. Optional Accessories Adjusting the Knife Holder with Anti-roll Guide You can adjust the height of the anti-roll guide system using the knurled nut ( 1 ): • If you turn the nut counterclockwise, the anti- roll guide system moves toward the blade. • If you turn the nut clockwise, the anti-roll guide syste[...]

59 Leica CM1950 – Cryostat Cleaning the knife holder CN Daily cleaning For daily cleaning, it is enough to fold the anti-roll guide system forwards and remove the section waste from the knife holder using a dry brush. Please use a cold brush, as otherwise the section waste will thaw and stick to the knife holder. It is not necessary to oil the pa[...]

60 Operating Manual V 1.1 – 05/2007 1 Knife holder CN with anti-roll guide – Moving the clamping chucks The large specimen disc (80 x 50 mm) should pref- erably be used with knife holder CN and the 16 cm C-profile steel knife. The clamping jaws are factory-installed in the knife holder with a clearance of 64 mm. If neces- sary, both clamping ch[...]

61 Leica CM1950 – Cryostat Knife holder CNZ with anti-roll guide • Pressure plate permits the full length of the knife to be utilized. • Use hard metal and steel knives here. Fig. 52 11. Optional Accessories The height of resharpened knives must be adjusted using knurled screws (3) (approx. 1 mm under the edge of the clamping jaw). Ensure tha[...]

62 Operating Manual V 1.1 – 05/2007 Fig. 56 Bacterial filter • Bactericidal filter 350/5865, pack of 1 unit. Rec- ommendation: replace bactericidal filter every three months. (note date on filter when installing it) Anti-roll guide system Anti-roll plate (with glass stage plate) Available with various spacers: • 70 mm - 50 μm, for section th[...]

63 Leica CM1950 – Cryostat Fig. 57 Fig. 58 Replacement filters for extraction system (pack of 25) • Change when vacuum power is impaired. • When using the cryostat frequently, check the filter bag daily and replace as required. Vacuum extraction system A - Hose B - Hose adapter, black (for the filter bag in the instrument) C - Hose adapter, w[...]

64 Operating Manual V 1.1 – 05/2007 Fig. 60 A Heat extractor with parking station, stationary • for installation in the cryostat - consisting of: support device, heat extractor and parking sta- tion B Heat and cold extractor • suitable for two different applications: extracts cold when stored in warm environment; ex- tracts heat, when stored [...]

65 Leica CM1950 – Cryostat Footswitch, for use with motorized instruments only Fig. 63 Fig. 66 Fig. 65 11. Optional Accessories Brush shelf, for use with blade holder CE Fig. 64 Adjustable support, individually height-adjustable foot rest with 5 set- tings. Storage system, movable for installation in front area of cryostat - for cold storage of p[...]

66 Operating Manual V 1.1 – 05/2007 12. Maintenance and Cleaning 1 Fig. 67 12.1 General Maintenance The microtome is virtually maintenance-free. To ensure a smooth operation of the instrument over several years we recommend the following: • At least once a year, have the instrument inspected by a qualified service engineer authorized by Leica. [...]

67 Leica CM1950 Cryostat 12. Maintenance and Cleaning Basically, we recommend UV disinfection (see page 35 for detailed instructions). For easy-to-use spray disinfection we recommend Leica Cryofect. (Cryofect is not available in all countries!) The cryostat has to be disinfected after each daily use. Comply with the instructions for use! The glass [...]

68 Operating Manual V 1.1 – 05/2007 12.3 Replacement of the Lamp/UVC Lamp Turn the instrument off with the automatic power fuse and pull the power 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. Beware of the metallic mercury in the UV lamp; h[...]

69 Leica CM1950 – Cryostat Installation of the new lamp Type: OSRAM DULUX L 18 W/ 840. • Hold the lamp in the correct mounting posi- tion as shown and push it to the left until it engages in the holder. • Slightly lift the sliding window ( 1 ) holding it by the grip ( 2 ) and pull it out to the front. 1 2 3 4 Removal of the lamp • The lamp [...]

70 Operating Manual V 1.1 – 05/2007 13. Decontamination Certificate (master) Dear Customer, Any product which is to be returned to Leica Microsystems or serviced on site, must be cleaned and decontaminated in the appropriate manner. Leica accepts that it is not possible to decontami- nate for CJD and its variants and in this instance following cu[...]

71 Leica CM1950 – Cryostat 13. Decontamination Certificate (master) Leica Biosystems Nussloch GmbH Heidelberger Str. 17-19 69226 Nussloch, Germany Phone: ++49 (0) 6224 143 0 Fax: ++49 (0) 6224 143 200 www.leica-microsystems.com Important - to avoid refusal of shipment: Place one copy in the unit prior to packaging or hand it over to the service e[...]

We herewith declare, in exclusive responsibility, that the Leica CM1950 Cryostat was developed, designed and manufactured to conform with the • Council Directive 2006/95/EC (Low Voltage), • Council Directive 89/336/EEC, Appendix I (Electromagnetic Compatibility) and • European council Directive 98/79/EC (IVD) including their amendments up to [...]

73 Leica CM1950 Cryostat 15. Warranty and Service Warranty Leica Biosystems Nussloch GmbH guarantees that the contractual product delivered has been subjected to a comprehensive quality control proce- dure based on the Leica in-house testing standards, and that the product is faultless and complies with all technical specifications and/or agreed ch[...]

74 Operating Manual V 1.1 – 05/2007 16. People's Republic of China[...]