Leica CM1950 Operating Manual
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Leica CM1950 Operating Manual

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Cryostat
Operating Manual
Leica CM1950 , Rev C, English, 1v3 – 04/2010
Always keep this manual near the instrument.
Read carefully prior to working with the instrument!

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Summary of Contents for Leica CM1950

  • Page 1 Cryostat Operating Manual Leica CM1950 , Rev C, English, 1v3 – 04/2010 Always keep this manual near the instrument. Read carefully prior to working with the instrument!
  • Page 3 Leica reserves the right to change technical spec- ing thorough investigation in this field. We are ifications as well as manufacturing processes under no obligation to update the present manu- without prior notice.

  • Page 4: Table Of Contents

    Table of contents Important Information ........................ 6 Symbols used in this manual and their meaning ..............6 Qualification of personnel ......................6 Designated use ..........................6 Instrument type ..........................6 Safety and Design ........................7 General safety instructions......................7 Unpacking and installation ......................7 Safety features ..........................
  • Page 5 Table of contents Instrument Controls ......................... 30 Control panel fields on the CM1950 – Control panel field 1 ..........30 Control panel field 2 – Electric coarse feed, sectioning and trimming thickness display ..32 Control panel 3 – Motorized sectioning (optional) ............... 34 Working with the Instrument ....................

  • Page 6: Important Information

    1.2 Qualification of personnel Instrument type The Leica CM1950 may be operated by trained All information given in this instruction manual laboratory personnel only. applies only to the instrument type indicated on All laboratory personnel designated to operate the title page.

  • Page 7: Safety And Design

    2. Safety and Design General safety instructions The CM1950 is a cryostat with an encapsulated microtome and separate spec- imen cooling. It is primarily used for work in the area of fast-cut diagnostics. The displays and instrument controls are easy to operate due to their largely self-explanatory symbols.
  • Page 8 Safety and Design 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.
  • Page 9 The rollers (14) must run over the cover hinges (2) at the front and rear. Danger of tipping! Fig. 6 1. Carefully roll the instrument backwards over the ramp from the pallet. 2. Push the instrument to the installation location on the castors (14). Leica CM1950 – Cryostat...

  • Page 10: Safety Features

    Safety and Design Safety features The instruction manual includes important information related to the operating safety and maintenance of the instrument The Operating Manual is an important part of the product, and must be read carefully before startup and use and must always be kept near the instrument. If additional requirements on accident prevention and environmental protection apply in the country of operation, this Operating Manual must be supplemented by appropriate instructions to ensure compliance with such requirements.

  • Page 11: Securing/Locking The Handwheel

    The handle automatically engages in this position. An important safety device on the cryostat is the centering of the handwheel for motorized instruments. Rotate the handwheel only if the refrigeration system is on and the cryochamber is cold. Fig. 9 Leica CM1950 – Cryostat...

  • Page 12: Cleaning, Disinfection - Turning The Instrument Back On

    It is not necessary to remove the microtome for disinfection. • The instrument has been designed for UV disinfection! Spray disinfec- tion with Leica Cryofect is also possible, thanks to the special insulation of the microtome. (Cryofect is not available in all countries!) Remove section waste after EVERY sectioning operation and BE- FORE changing specimens.

  • Page 13: Handling Specimens - Defrosting

    Technical Ser- vice. If any metallic mercury is released, handle it carefully and dispose of it properly. If both disinfection indicator lights are blinking alternately, the UV tube must be replaced! Leica CM1950 – Cryostat...

  • Page 14: Technical Data

    Technical Data All specifications related to temperature are valid only up to an ambient temperature of 18 °C to 35 °C and a relative humidity of no more than 60%. Instrument type Nominal voltage (±10 %) 100 V AC 120 V AC 230 V AC Nominal frequency 50/60 Hz...
  • Page 15 3. Technical Data Refrigeration system CM1950 , 50 Hz CM1950 , 50 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...

  • Page 16: Standard Delivery

    Compare the delivered components with the parts list and your order. Should you find any discrepancies, please contact your Leica sales office without delay. A choice of different knife holders is available for the CM1950 . Operating Manual, Rev C, 1v3 – 04/2010...
  • Page 17 Compare the delivered components with the parts list and your order. Should you find any discrepancies, please contact your Leica sales office without delay. A choice of different knife holders is available for the CM1950 . Leica CM1950 – Cryostat...

  • Page 18: General Overview

    General Overview Emergency-stop On/off switch, also circuit switch (motorized breaker instruments only) Footswitch dummy Cryostat chamber (motorized (with activated instruments only) UV disinfection) Handwheel in 12 o'clock position Bacterial filter (optional, only for instruments with filter) Condenser Catch tank Unscrew and align for condensate adjusting feet after transport...

  • Page 19: Control Panel Fields And Cryostat Chamber

    (the coarse filter insert is behind it) 9 - Shelf, movable (optional) 15 - Object head, directional 10 - Blade holder CE with blade ejector (a) 16 - Waste tray 10b- Finger rest on the blade holder CE Leica CM1950 – Cryostat...

  • Page 20: Installation

    Installation 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. •...

  • Page 21: Transport To The Site

    (Refer to p. 26.) Fig. 12 When not using the extraction for a long period, tightly close the opening for the extraction hose using the silicone stop- per (E, page 28) included in the scope of 2 (covered) delivery. Leica CM1950 – Cryostat...

  • Page 22: Assembling The Handwheel

    Installation Transport with a fork lift • The instrument can be transported with a fork lift. To ensure a safe transport 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.

  • Page 23: Locking/Unlocking The Handwheel

    (see page 16, General overview) if no footswitch (optional) is used. If the red LED in the E-STOP field in control panel field 3 is illuminated, either: • emergency-stop function active, or • footswitch dummy (optional footswitch) connected incorrectly or not at all. Fig. 18 Leica CM1950 – Cryostat...

  • Page 24: Connection To Power Supply System

    Installation 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 during this time due to temperature fluctuations must be allowed to dry completely.

  • Page 25: Installing Accessories/Inserting Chamber Accessories

    (height). • Once installed, the user can adjust the height of the footrest at any time by relocating it (3) to the desired height on both sides in the holder (2). Fig. 20 Leica CM1950 – Cryostat...

  • Page 26: Installing The Storage Systems (Optional)

    Installation 6.5.2 Installing the storage systems (optional) reasons accessibility, the (optional) storage system must always be installed first. To do so, remove the insert (1), place the frame (2) in front of the bore and tighten the screws/ washers on the cryostat housing using the No. 4 Allen key.

  • Page 27: Installing The Heat Extractor, Stationary (Optional)

    When removing the knife holder base (2) from the refrigerated cryostat chamber, hold it by the grip points (a—front and rear) to keep your fingers from freezing. Safety gloves must be worn! Fig. 25 Leica CM1950 – Cryostat...

  • Page 28: Inserting/Changing The Bacteria Filter

    Installation 6.5.7 Inserting/changing the bacteria filter • To change the filter, follow the opposite proce- dure: press the filter to the right, then pull it to the left and out of the tube. • The filter must be changed approx. every 3 months (we recommend writing the date on the filter using a marker).

  • Page 29: Installing The Section Extraction (Optional) - Use With Blade Holder Ce Only

    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 alcohol. After several cleanings, the hose must be replaced (see order information on p. 49)! Leica CM1950 – Cryostat...

  • Page 30: Instrument Controls

    Instrument Controls Control panel fields on the CM1950 – Control panel field 1 Display of actual and target chamber Display of actual and target specimen Display of real time, defrost time temperatures head temperatures and error messages. Fig. 32 Button to...
  • Page 31 When the condenser starts up, the flashing starts and the 10 minutes are counted down. (For exact instructions on using the chamber, specimen head and real time display fields, refer to the chapter on "Working with the Instrument" on page 37 of this Operating Manual.) Leica CM1950 – Cryostat...

  • Page 32: Control Panel Field 2 - Electric Coarse Feed, Sectioning And Trimming Thickness Display

    Instrument Controls Control panel field 2 – Electric coarse feed, sectioning and trimming thickness display Illuminates in For trimming values above a sec- yellow when the tion thickness of 200 µm, the specimen is in LED for trim display flashes to expressly retraction.
  • Page 33 • For sectioning, turn the handwheel a short distance (approx. 1/4 turn) forwards and back (rocking mode)—only possible at rear (handwheel in approx. 12 - 3 o'clock position). Every change in rotation direction is electronically detected and automati- cally translated into a specimen feed or retraction movement. Leica CM1950 – Cryostat...

  • Page 34: Control Panel 3 - Motorized Sectioning (Optional)

    Instrument Controls Control panel 3 – Motorized sectioning (optional) Switching the sectioning If emergency stop has been activated, mode ("CUT MODE") from the cutting mode must be selected continuous stroke ("CONT") again. to single stroke ("SINGLE") (active) The red LED in the E-STOP field indicates either: Motor speed controller (0-100%) •...
  • Page 35 UVC disinfection is effective when disinfecting surfaces and air within the irradiated working area of the cryostats Leica CM1850UV, CM1900UV and CM1950 at -20°C (Table 1). For powerful disinfec- tion, we recommend irradiation for three hours (CM1850UV/CM1950) and four hours (CM1900UV).

  • Page 36: Working With The Instrument

    Working with the Instrument 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 applicable, specimen discs into the cryostat chamber.

  • Page 37: Configuring The Parameters

    Refer to page 52 for a table with guide values. The temperature values given there are based on experience, but are intended sole- ly as guide values, as any tissue may require particular adjust- ments. Leica CM1950 – Cryostat...
  • Page 38 Working with the Instrument Programming the specimen temperature • Select the desired temperature of the specimen. • The specimen temperature is set and indicated on the control panel marked with the specimen head symbol. The actual temperature is the standard indication. Briefly pressing the "+"...
  • Page 39 After defrosting the freeze shelf, carefully wipe it out, as otherwise a lot of water collects in the channel. Ice does not melt during nor- mal defrosting. Leica CM1950 – Cryostat...
  • Page 40 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. •...
  • Page 41 • You can switch by pressing the "+" or "-" button. • "Retraction on" means a specimen retraction of 20 µm in manual mode. During motorized sectioning, the retraction value is speed-depen- dent and cannot be changed by the user. Leica CM1950 – Cryostat...

  • Page 42: Working With The Precooled Cryostat

    Working with the Instrument 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 specimen.

  • Page 43: Trimming With Extraction - Anti-Roll Guide Installed

    • 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. Leica CM1950 – Cryostat...
  • Page 44 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 again to disable it.

  • Page 45: Cutting With Extraction - Anti-Roll Guide Installed

    After removing the section, wipe off moisture/condensate from the pressure plate–otherwise, the next sections will become jammed. Sectioning with extraction without anti-roll guide (brush tech- nique) is not possible, as the position of the pressure plate means that no suitable air flow is obtained. Leica CM1950 – Cryostat...
  • Page 46 Working with the Instrument 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.
  • Page 47 • Check the bacterial filter (in the front of the unit, see page 28), change it at least every 3 months. • Move the anti-roll guide to the side. • Spray the cryostat chamber using Cryofect. • Activate the UVC disinfection. Leica CM1950 – Cryostat...

  • Page 48: Troubleshooting

    Troubleshooting Problems during work Problem Causes Remedies Frost on chamber walls and Cryostat is exposed to air currents Change place of installation for the microtome (open windows and doors, air condi- cryostat. tioning). Frost built-up by breathing into the Wear mouth protection. cryochamber.
  • Page 49 Check knife clamping. Knife not clamped tightly enough. Check disc clamping. Specimen disc not clamped tightly enough. Apply cryocompound to warm disc, Cryocompound applied to cold mount specimen and freeze. specimen disc; specimen detached from disc after freezing. Leica CM1950 – Cryostat...
  • Page 50 9. Troubleshooting Problem Causes Remedies Blunt knife. Use different part of the knife. Inappropriate section thickness. Select correct section thickness. Incorrect clearance angle. Set correct angle. Microtome not dried thoroughly Dry microtome. enough. Dried specimen. Prepare new specimen. Anti-roll plate is too warm or incorrect- Cool down anti-roll plate, or Tissue sticks to the anti-roll ly positioned.
  • Page 51 UV radiation provided by UV tube no Replace UV tube following the alternately longer sufficient. manufacturer’s instructions. Image of an open-ended wrench Contact technical service and appears due to fault to be rectified. follow the instructions given. Leica CM1950 – Cryostat...

  • Page 52: Temperature Selection Chart (In Minus °C)

    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...

  • Page 53: Optional Accessories

    Knife support CN ................14 0477 42370 Knife holder attachment CNZ, assy..........14 0477 42363 Anti-roll plate assembly glass 50mm ........... 14 0419 33981 Heat and cold extractor, assy............14 0477 41039 Cold extractor, assy............... 14 0477 43737 Leica CM1950 – Cryostat...
  • Page 54 Hose set, 5 pieces ................14 0477 44469 Filter assembly 25 pieces .............. 14 0477 44307 VPE Leica Cryofect, 4x 350 ml ............14 0387 42801 Safety gloves size M ..............14 0340 29011 Safety gloves size S ............... 14 0340 40859 Footswitch, dummy plug CM 3050 ..........
  • Page 55 4. Screw the knurled nut (13) from below onto the shaft (11). Fig. 42 The glass of the anti-roll guide can be used from all 4 sides when replace- ment is necessary (the glass stage plate can be reordered). Leica CM1950 – Cryostat...
  • Page 56 11. Optional Accessories The anti-roll guide and Red elements on the blade or knife pressure plate must be holders, such as the knife guard and parallel to each other. ejector, are protective devices that may not be removed. • Blade ejector (1) and knife guard (2) •...
  • Page 57 The safety gloves included in the stan- dard scope of delivery must be worn when disposing of the blade! Once the blade has been removed from the blade holder, it is disposed of into the dispenser container (underside, see image). Fig. 45 Leica CM1950 – Cryostat...
  • Page 58 11. Optional Accessories Lateral shift for Blade Holder CE If the sectioning results are not satisfactory, the knife holder can be shifted sideways in order to use another part of the blade. To do so, follow these steps: 1. Release the clamping lever (8) by folding it back and move the knife holder back until the desired position is reached (3-point click stop enables you to accurately find a new cutting...
  • Page 59 Generally, we recommend pre-adjusting the anti-roll guide system at a high section thickness (e.g. 10 µm). Start from there and work your way down to the desired section thickness in small increments, readjusting the anti-roll guide system at each increment using the knurled nut. Leica CM1950 – Cryostat...
  • Page 60 (with alcohol or acetone). For disinfection, commercially avail- able, mild cleaners and disinfectants Fig. 49 can be used—we recommend Leica Cryofect. 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.
  • Page 61 (3a). Never work with only one clamping jaw, as this does not ensure the stability required for the sectioning process. Also, a long knife will not be sufficiently protected by the knife guard in this instance. Leica CM1950 – Cryostat...
  • Page 62 11. Optional Accessories Knife guard/lateral movement for knife holder CN The knife guard (18) is fixed and integrated into the clamping jaws. The knife guard has handles (19) that allow it to be moved. The knife guard is adequate for knives up to 22 cm in length. Always cover exposed parts of the knife blade after sectioning.
  • Page 63 3 months. (Write the installation date on the filter) Filter bags and bacteria filter must be disposed of according to applicable lab- oratory regulations for infectious mate- rial. Filters must be replaced, not cleaned. Fig. 57 Leica CM1950 – Cryostat...
  • Page 64 11. Optional Accessories Replacement filter for extraction system, pack of 25 • Change when extraction power is impaired. • When using the cryostat frequently, check the filter bag daily and replace as required. Fig. 58 Vacuum extraction system A - Hose B - Hose adapter, black (for the filter bag in the instrument) C - Hose adapter, white (for suction nozzle D...
  • Page 65 28 x 28 mm Medium shelf with 4 recesses L x W x H: 24 x 24 x 6 mm 37x 37 mm Small shelf with 4 recesses L x W x H: 18 x 18 x 6 mm Leica CM1950 – Cryostat...
  • Page 66 11. Optional Accessories Footswitch, for use with motorized instruments only. The foot switch can be used to control the motorized sectioning process. It also has a function that is similar to the emergency stop function. Caution! Fig. 64 In addition to the foot switch, all control panel functions and all buttons on the instrument continue to be active.
  • Page 67 5 adjustment options. Fig. 65 Brush shelf, for use with Blade Holder CE Fig. 66 Storage system, movable for installation in the front part of the cryostat for cooled storage of preparation aids Fig. 67 Leica CM1950 – Cryostat...

  • Page 68: Maintenance And Cleaning

    Do not use organic solvents or any other aggressive substances for cleaning and disinfecting! Use only disinfectants listed in this manual such as Leica Cryofect (alcohol or other common alcohol-based disinfectants). • Drain the cleaning liquid through the hose after the prescribed reagent time is over and collect it in the waste container (1).
  • Page 69 Fig. 68 Basically, we recommend UV disinfection (see page 35 for detailed instruc- tions). 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 anti-roll plate can remain in place during disinfection.

  • Page 70: Changing Fuses

    12. Maintenance and Cleaning 12.2 Changing fuses • In case of power supply faults, please contact an authorized Leica service technician immediately. Do not carry out any repairs on your own as this will invalidate the warranty. Repairs may only be carried out by qualified service engineers au- thorized by Leica.
  • Page 71 30 seconds, the running-time meter for Fig. 72 the UVC lamp is reset. This is required every time the UVC lamp is replaced to Dispose of the UVC lamp separately! ensure sufficient power for disinfection! Leica CM1950 – Cryostat...

  • Page 72: Replacing The Fluorescent Lamp

    12. Maintenance and Cleaning 12.3.1 Replacing the fluorescent lamp Switch the instrument off and discon- nect the power plug before replacing the fluorescent lamp! If the lamp is broken, it must be replaced by the technical service, as the replace- ment involves a high risk of injury.

  • Page 73: Decontamination Certificate (Master)

    (master) Dear Customer, Any product which is to be returned to Leica Biosystems or serviced on site, must be cleaned and decon- taminated in the appropriate manner. Since it is not possible to decontaminate against diseases caused by prions such as CJD, BSE or CWD, instruments that have come into contact with prion-contaminated spec- imens cannot be returned to Leica Biosystems for repairs.
  • Page 74 Important - to avoid refusal of shipment: Place one copy in the unit prior to packaging or hand it over to the service engineer. Customer assumes all responsibility for the immediate return shipment of articles sent to Leica without proper decontam- ination documentation.

  • Page 75: Warranty And Service

    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 char- acteristics warranted.

  • Page 76: Peoples Republic Of China

    Peoples Republic of China Operating Manual, Rev C, 1v3– 04/2010...

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