Leica CM3050 S manuale d’uso

Leica CM3050 S Cryostat Instruction Manual Leica CM 3050 S – Cryostat English V1.2– 11/2006 Always keep this manual near the instrument. Read carefully prior to operating the instrument.[...]

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3 Leica CM 3050 S – Cryostat Issued by: Leica Microsystems Nussloch GmbH Heidelberger Str. 17 - 19 D-69226 Nussloch Germany Phone: +49 6224 143-0 Fax: +49 6224 143-200 Internet: http://www.histo-solutions.com Serial No.: .............. ...................................................... Year of Manufacture: ....................................[...]

4 Instruction Manual V1.2 - 11/2006 Table of Contents 1. Important information ........................................................................................................ ........ 7 1.1 Symbols used in this manual and ............................................................................................. ....... 7 1.2 Designated [...]

5 Leica CM 3050 S – Cryostat Table of Contents 4. Instrument Properties ........................................................................................................ ...... 21 4.1 Overv iew ............................................................................................................................................ 21 4.2[...]

6 Instruction Manual V1.2 - 11/2006 Table of contents 6. Troubleshooting, applications tips ....................................................................................... 47 6.1 Displayed error messages .................................................................................................... .......... 47 6.1.1 Other error indic[...]

7 Leica CM 3050 S – Cryostat The chapters of the Leica CM3050 S cryostat instruction manual: Chapter 1 Structure of the manual : • Table of contents • Important information on this manual Chapter 2 Safety • Read this chapter before operating the instrument! Chapter 3 Installation • Unpacking and installation • Standard delivery, informa[...]

8 Instruction Manual V1.2 - 11/2006 General This instruction manual includes important instructions and information related to the operating safety and maintenance of the instrument. The instruction manual is an important part of the product. It must be read carefully before using the instrument for the first time and must always be kept with the i[...]

9 Leica CM 3050 S – Cryostat 2.1 General information on instrument design and safe handling This instrument has been built and tested in ac- cordance with the following safety regulations on electrical measuring, control, regulating and laboratory devices: • DIN EN 292 • DIN EN 61010-1 • EN 50082-1 • EN 55011 • IEC 1000-4 as well as acc[...]

10 Instruction Manual V1.2 - 11/2006 Instruments with sectioning motor: The sectioning motor is now blocked. All instruments: The message ‘LOCKED’ in the display of control panel 1 indicates that the handwheel has been locked: Unlocking: • To unlock, push locking pin ( 2 ) to the left into position ( 2a ). • Display indication ‘LOCKED’ [...]

11 Leica CM 3050 S – Cryostat 2.2.3 Emergency stop function (instruments with sectioning motor only) The emergency stop is activated via the red emergency stop button in control panel 2 or via the footswitch Activating the emergency stop function • Press emergency stop button or step on footswitch forcefully. --> STOP (red) lights up. As soo[...]

12 Instruction Manual V1.2 - 11/2006 2. Safety 2.3 Safety instructions: handling and operation of the instrument 2.3.1 Transport • To avoid severe damage to the instrument by running it while the compressor oil is dis- placed from its regular position: • Do not tilt the instrument, only transport in an upright position. • After transport, wai[...]

13 Leica CM 3050 S – Cryostat 2.3.4 Handling microtome knives/blades • Danger! --> Microtome knives and disposable blades have extremely sharp cutting edges and can cause serious injuries. Therefore: • Handle knives / blades with utmost care. • Never leave any knives / blades in unpro- tected places! • Never place a knife, no matter wh[...]

14 Instruction Manual V1.2 - 11/2006 2. Safety 2.3.9 Infectious/radioactive material • Danger! Use caution when working with potentially infectious specimens: -->Risk of infection! • When working with potentially infectious / radioactive specimens: • Wear protective clothes (gloves, protec- tive boots, mask, lab coat), in compliance with r[...]

15 Leica CM 3050 S – Cryostat 2. Safety 2.3.11 Removing/reinstalling the microtome • Before removing the microtome: • Switch instrument off. • Unplug from mains. • Place handwheel grip in lowest position and lock. --> When removing the microtome, the specimen head must always be lok- ked in the lowest position. Otherwise the upper part[...]

16 Instruction Manual V1.2 - 11/2006 3.2 Unpacking and installation Unpacking instructions are always located in a transparent protective envelope on the outside of the instrument shipping crate. Make sure to read and follow all safety instructions provided in chapter 2.3.1 ‘Transport’ and on the unpacking instructions! 3.2.1 Repacking We recom[...]

17 Leica CM 3050 S – Cryostat 3. Installation 3.3 Standard delivery 1 Cryostat with microtome ............................................................................. 0460 32037 1 Handwheel with marking, antibacterial ................................................... 0471 42558 1 Heat extractor, stationary .................................[...]

18 Instruction Manual V1.2 - 11/2006 Further accessories: Further accessories which you ordered will be included in the box containing the standard delivery items. Knife holders are delivered with anti-roll guide, knife guard, and a separate instruction manual. In case of non-configured instruments, the knife holder is not a part of standard delive[...]

19 Leica CM 3050 S – Cryostat 3.4 Installing/inserting accessories 3.4.1 Installing the handwheel • Insert pin ( 1 ) of the handwheel shaft into hole ( 2 ). • Place spring washer ( 3 ) onto screw ( 4 ) as shown. • Tighten screw ( 4 ) with an Allen key. • Cover with selfadhesive disc ( 5 ). For purposes of transport (e.g. narrow doors), th[...]

20 Instruction Manual V1.2 - 11/2006 3.4.3 The footswitch Function The footswitch performs the same task as the RUN/STOP and RUN/ENABLE keys (activating/ deactivating motorized sectioning / trimming). In addition, the footswitch can be used to acti- vate the emergency stop function. Models with footswitch: All instruments with sectioning motor. Imp[...]

21 Leica CM 3050 S – Cryostat 4. Instrument Properties 4.1 Overview 1 Cryostat CM3050 S 2 Control panel 1 3 Control panel 2 4 Storage shelf, left 5 Storage shelf, right 6 Rubber mat 7 Mains switch / Automatic cutout for instrument, Automatic cutout for sectioning motor, Footswitch port 8 Quick-freeze shelf 9 Stationary heat extractor 10 Mobile he[...]

22 Instruction Manual V1.2 - 11/2006 4. Instrument Properties 15 13 14 11 17 11 21 20 19 18 16[...]

23 Leica CM 3050 S – Cryostat 4. Instrument properties Operating temperature range (ambient temperature): + 18°C to + 40°C. All specifications related to tempe- rature are valid only up to an ambient temperature of + 22°C and relative air humidity of less than 60 %! Refrigeration system 50 Hz 60 Hz Cryochamber Temperature selection range 0° C[...]

24 Instruction Manual V1.2 - 11/2006 4. Instrument properties Microtome Rotary microtome Section thickness setting 0.5 - 300 μm Overall specimen feed 25 mm Vertical stroke 59 mm Specimen retraction 50 μm Maximum specimen size 40 x 55 mm Specimen orientation 8° (x-, y-, z-axis) Trimming (6 thickness settings) 5 - 150 μm Motorized coarse feed - s[...]

25 Leica CM 3050 S – Cryostat 5. Operation 5.1 Operating the instrument for the first time Prior to setting up the instrument, make sure to read the instruction manual carefully and in full. When setting up the instrument, please follow the individual chapters in the following order: • First, study chapters 1, 2, and 3 - install the instrument [...]

26 Instruction Manual V1.2 - 11/2006 5.2 Switching on/fuses Switching on • To switch on the instrument, place switch ( 1 ) in upper lock-in position. Mains switch / automatic cutout for the instrument: In addition to its function as ON/OFF switch, switch ( 1 ) is also the automatic cutout for the in- strument. If the automatic cutout has been tri[...]

27 Leica CM 3050 S – Cryostat 5.3.1 Key functions in control panel 1 5.3 Control panel 1 ON/OFF key for cryochamber illumination. To select the menu items for setting / changing instrument parameters. In the status indication, pressing the ar- row keys displays the individual current parameters. In the menu, pressing the arrow keys changes set va[...]

28 Instruction Manual V1.2 - 11/2006 5. Operation Meaning of indication: Operator activity: yes no no Acknowledge indication by pressing once briefly. Initialization phase 1 Initialization phase 2 Software version Selected display language Sectioning motor speed level ‘slow’ / (fast) --- --- --- Select language via keys Select speed level via k[...]

29 Leica CM 3050 S – Cryostat 5. Operation Once the instrument has gone through the initial sequence of indications, you can go on to the menu functions to select and/or change instru- ment parameters. If display is locked: Press key until display background lights up. Next, press the menu key. With each key stroke the following parameters will b[...]

30 Instruction Manual V1.2 - 11/2006 5. Operation 2. Cryochamber temperature: Press key until menu item ‘Set chamber temperature’ is displayed: --> Continue as described for specimen temperature. 3. Preset counter (instruments with sectioning motor only): Press key until menu item ‘Preset counter’ is displayed: = Preset counter activated[...]

31 Leica CM 3050 S – Cryostat The automatic defrost cycle takes place once every 24 hours at the selected defrost time. 7. Duration of defrost cycle: Press key until menu item ‘Defrost period’ is displayed: • Use defrost keys ( or ) to adjust de- frost time. (Only if necessary - see below). Important information for setting the defrost para[...]

32 Instruction Manual V1.2 - 11/2006 5. Operation 5.3.5 Status display Each individual parameter is displayed in a certain area of the display screen. Digits 1 and 2 indicate sectioning stroke position: • Sectioning stroke (outside sectioning window) •  : Retraction stroke • [ ] : Sectioning stroke (inside sectioning window) Digits 3 - 6 i[...]

33 Leica CM 3050 S – Cryostat 5. Operation These parameters can be displayed via the ar- row keys. When pressing , the parameters are dis- played in the sequence listed on this page; when pressing , they are displayed in re- verse order. The display can be scrolled contin- uously, i.e. after the last parameter the display will automatically start[...]

34 Instruction Manual V1.2 - 11/2006 5. Operation 5.4 Control panel 2 Functional area A: • Sectioning modes • Sectioning window • Sectioning speed • Emergency stop function Functional area B: • Coarse feed • Trimming • Section thickness selection Functional area C: • Start / Stop sectioning Instruments without sectioning motor Instr[...]

35 Leica CM 3050 S – Cryostat Start / stop motorized sectioning Sectioning mode 1 - Intermittent stroke: • Simultaneously press and hold RUN/STOP and RUN/ENABLE or depress and hold footswitch. • To stop the sectioning stroke, release the keys or the footswitch: The specimen head stops immediately. Sectioning mode 2 - Continuous stroke: • Si[...]

36 Instruction Manual V1.2 - 11/2006 The value has been stored when the LED in the sectioning window key lights up briefly. If the LED flashes, the value has not yet been stored, because the user has tried to set a win- dow margin outside the possible window range (the area in which the sectioning window can be set is slightly smaller than the tota[...]

37 Leica CM 3050 S – Cryostat 5.4.3 Selecting sectioning speed • Sectioning speed is selected via the sliding potentiometer. Speed can be changed while sectioning is in motion. Caution when working in continuous mode: If the slider is set to ‘0’, the sectioning motion stops, however, the actual sectioning mode is still active - just operati[...]

38 Instruction Manual V1.2 - 11/2006 5. Operation 5.4.5 Coarse feed keys The four coarse feed keys in functional area B function as follows: Coarse feed ‘fast backward’ - Travelling speed 1,000 μm/s • Press key to activate. Specimen head travels backward (= away from knife): a. until it reaches the rear limit position, b. until it is stopped[...]

39 Leica CM 3050 S – Cryostat 5. Operation Press this key to alternate between the two functions. illuminated: Trimming mode is active off: Sectioning mode is active. Selecting trimming thickness • Activate trimming mode • Select trimming thickness (= sectioning thickness at which trimming is carried out) by pressing or . The selected thickne[...]

40 Instruction Manual V1.2 - 11/2006 5.5 Daily operation of the instrument • Select chamber temperature and, in instru- ments with specimen cooling, specimen temperature according to the type of speci- men to be sectioned (see also chapter 6.3 ‘Reference chart for temperature settings’). • Do not continue with step 5.5.1 before the actual c[...]

41 Leica CM 3050 S – Cryostat 5.5.2 Freezing specimens onto specimen In instruments with specimen cooling, the 90 ° prism, which is inserted into the specimen head, is an additional option to freeze specimens at maximum temperature . Installing the 90 ° prism • Loosen screw (3) at specimen head. • Insert pin (1) of prism completely into rec[...]

42 Instruction Manual V1.2 - 11/2006 5. Operation 5.5.4 Inserting specimen discs into the • Lock handwheel in upper position. If a knife has already been inserted into the knife holder, cover knife edge with knife guard! • Loosen clamping screw (3). • Insert specimen disc in opening (2) (rear surface of specimen disc must fit tightly against [...]

43 Leica CM 3050 S – Cryostat 5. Operation Inserting the knife • Insert the knife / disposable blade into the knife holder. • Select appropriate clearance angle at knife holder (settings between 4° and 6° are suit- able for the majority of applications). For details on how to insert and ad- just the knives, please refer to the separate inst[...]

44 Instruction Manual V1.2 - 11/2006 5. Operation 5.6 Finishing work 5.6.1 Daily routine steps Remove all specimens from the cry- ochamber and store safely! S e e also corresponding safety instruc- tions in chapter 2.3.7 ‘Defrosting / handling frozen tissue’! • Lock handwheel. • Remove knife from knife holder, disinfect, dry (see chapter 7 [...]

45 Leica CM 3050 S – Cryostat  HO ME   5.7 Defrosting The Leica CM3050 S has 3 (instruments without specimen cooling: 2) different defrosting options. - Automatic chamber defrost cycle - Manual chamber defrost cycle - Specimen head defrosting (manual) - instru- ments with specimen cooling only. Read the safety instructions in chap- te[...]

46 Instruction Manual V1.2 - 11/2006 SE T T E M P OT - 3 5 ° C   SE T T E M P C T - 3 0 ° C  - 5. Operation 5.7.2 Manual chamber defrost cycle In addition to the automatic defrost cycle, a manual chamber defrost cycle can be activat- ed when necessary. • Press until menu item ‘Set chamber temperature’ is displayed: • Press . A br[...]

47 Leica CM 3050 S – Cryostat 6. Troubleshooting, applications tips 6.1 Displayed error messages The autodiagnosis function of the CM3050 S displays error messages in case of certain mal- functions. 1. Power failure This error message is displayed after the instru- ment had been disconnected from mains, either due to power failure or because the [...]

48 Instruction Manual V1.1 02/2000 6. Troubleshooting, applications tips 6.2 Potential problems - causes and remedies Remedies - Eliminate draft sources or change place of installation of the cryostat. - Close sliding window. - Consider wearing a mask. - Select lower temperature. - Wait until knife and/or anti-roll plate have reached chamber temper[...]

49 Leica CM 3050 S – Cryostat 6. Troubleshooting, applications tips Problem Sections curl on anti-roll plate. Scraping noise during section- ing stroke and specimen return stroke. Ondulated sections. Chatter marks form during sectioning. Condensation forms on anti-roll plate and knife during cleaning. Causes - Anti-roll plate does not pro- trude [...]

50 Instruction Manual V1.1 02/2000 6. Troubleshooting, applications tips Causes - Anti-roll plate protrudes too far beyond the knife edge. Adjust- ment was done in the direction of the knife. - Wrong temperature for the tissue to be sectioned. - Knife profile inappropriate for the specimen to be sectioned. - Ice build-up on the knife back. - Handwh[...]

51 Leica CM 3050 S – Cryostat 6. Troubleshooting, applications tips Problem Sections tear. Inconsistent or insufficient specimen feed. Specimen disc cannot be removed. Cryostat inoperational. Sectioning motor inoperational. No or insufficient refrigerating performance. Causes - Temperature too low for the tissue to be sectioned. - Knife blunt, di[...]

52 Instruction Manual V1.1 02/2000 6. Troubleshooting, applications tips 6.3 Reference chart for temperature settings (in minus °C) Tissue 10 - 15 15 - 25 25 - 50 Adrenal  Bone marrow ❄ Brain  Bladder  Breast - fatty  Breast - little fat  Cartilage  Cervical  Fatty  Heart and vascular  Intestinal  Kidney  Lary[...]

53 Leica CM 3050 S – Cryostat 7. Disinfection, cleaning and maintenance 7.1 Safety instructions When disinfecting the instrument, make sure to observe the following safety instructions: • ‘Disinfection and Cleaning’ (Chapter 2.3.10) • ‘Handling microtome knives / blades’ (Chapter 2.3.4) • ‘Defrosting / handling frozen tissue’ (C[...]

54 Instruction Manual V1.2 - 11/2006 7 Disinfection, cleaning and maintenance 7.3 Disinfection with conventional • Switch the instrument off. • Carefully remove knife / blade as well as spec- imens and accessories from the chamber. • Remove section waste from the chamber and dispose of it. • Remove sliding window (see chapter 7.6 ‘Ex- cha[...]

55 Leica CM 3050 S – Cryostat 7.5 Removing / reinstalling the microtome 7.5.1 Removing the microtome When removing the microtome make sure to observe the following safety instructions: • ‘Removing / reinstalling the microtome’ (Chapter 2.3.11) • ‘Defrosting / handling frozen tissue’ (Chapter 2.3.7) • ‘Infectious / radioactive mate[...]

56 Instruction Manual V1.2 - 11/2006 7.5.3 Reinstalling the microtome When reinstalling the microtome make sure to observe the following safety instructions: • ‘Removing / reinstalling the microtome’ (Chapter 2.3.11) • Display message ‘Dry Microtome’ (Chapter 2.3.12) • Lock handwheel in lower position. • Place microtome in chamber. [...]

57 Leica CM 3050 S – Cryostat Removal • The lamp (3) is located behind the glare shield (4). • Carefully hold lamp with both of your hands. Slightly pull the lamp downward at the left side to remove it from fastening clip (5). • Pull the lamp towards the left to remove it from its socket (6). Installing the replacement lamp • Hold the lam[...]

58 Instruction Manual V1.2 - 11/2006 8. Warranty and service Warranty Leica Microsystems Nussloch GmbH guarantees that the contractual prod- uct delivered has been subjected to a comprehensive quality control procedure based on the Leica in-house testing standards, and that the prod- uct is faultless and complies with all technical specifications a[...]

59 Leica CM 3050 S – Cryostat An electrohydraulic lifting device for individual working height adjustment is available as an optional accessory for the CM3050 S cryostat series. 1.1 Designated use The electrohydraulic height adjustment device has been designed exclusively for adjustment of the instrument working level to the individual height of [...]

60 Instruction Manual V1.2 - 11/2006 9. Appendix 1.4 Installation • Open the shipping box and remove all packing material including the Sty- rofoam pieces. • Take the height adjustment device out of the box. • At the place of installation, place the U-frame on the floor with the open side facing yourself. Adjust the U-frame parallel to the wa[...]

61 Leica CM 3050 S – Cryostat The control keys are located on top of the con- trol unit (see photograph). • To lift the instrument, press . • To lower the instrument, press . As long as a button is held, the instrument will go up or down until reaching the upper or lower limit. 1.6 Maintenance and care • Protect the lifting pistons in the f[...]

62 Instruction Manual V1.2 - 11/2006 2.1 Knife holders Knife-holder CN for reusable microtome knives´w/anti-roll guide, adjustable knife guard and knife support bars for slim and wide knives ............................................. 0419 18649 Knife support bar for knife holder CN, for slim knives ..............................................[...]

. . . . . . . . . . . . . . . . . . . . . . . . . . . . Dag Graupner Managing Director We herewith declare, in exclusive responsibility, that the Leica CM3050 S – Cryostat was developed, designed and manufactured to conform with the • Council Directive 73/23/EEC (Low Voltage), • Council Directive 89/336/EEC, Appendix I (Electromagnetic Compat[...]

64 Instruction Manual V1.2 - 11/2006 11. Decontamination Certificate (master) Dear Customer, any product which is be returned to Leica Microsystems or serviced on site, must be cleaned and decontaminated in the appropriate manner. This certificate, duly completed, must be placed in the unit prior to packaging or handed over to the service engineer.[...]

65 Leica CM 3050 S – Cryostat 11. Decontamination Certificate (master) Important - to avoid refusal of shipment: Place one copy in the unit prior to packaging or hand it over to the service engineer. Customer as- sumes all responsibility for the immediate return shipment of articles sent to Leica without proper decontamination documentation. If y[...]

66 Instruction Manual V1.2 - 11/2006[...]