Leica EM GP2 Instructions For Use Manual
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Leica EM GP2 Instructions For Use Manual

Leica EM GP2 Instructions For Use Manual

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From Eye to Insight
From Eye to Insight
EM GP2
EM GP2
Instructions for use
Instructions for use
Leica Microsystems CMS GmbH, Instructions for use, Revision 10/17
Leica Microsystems CMS GmbH, Instructions for use, Revision 11/17

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  • Page 1 From Eye to Insight From Eye to Insight EM GP2 EM GP2 Instructions for use Instructions for use Leica Microsystems CMS GmbH, Instructions for use, Revision 10/17 Leica Microsystems CMS GmbH, Instructions for use, Revision 11/17...
  • Page 2 Important Note 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 to provide our customers with excellent products.
  • Page 3 EM GP2 Operating Manual Leica EM RAPID Operating Manual Operating Manual Leica EM GP2 Serial Number: Leica EM RAPID Serial Number: Date of purchase: Date of purchase: For the instrument serial number, please refer to the name type label on the...
  • Page 4 Leica reserves the right to change technical specifications and manufacturing processes without prior notice. Only in this way, Leica can continuously improve the technology and techniques used to provide its customers with excellent products.

  • Page 5: Table Of Contents

    Contents Table of Contents Introduction ............7 Installation ............20 1.1 Product Identification ........7 4.1 Work Area Conditions ........20 1.2 Field of Application and Intended Use ..7 4.2 Unpacking ............20 1.3 Qualification of Operating Personnel ...7 4.3 Assembly ............21 1.4 Symbols and Warnings ........8 4.4 Accessories ...........22 1.5 General Safety Information ......9 Operation ............23...
  • Page 6 Content 5.5 Freezing a Grid ..........37 5.5.1 Attaching the Forceps/Grid ......37 5.5.2 Lower Chamber ..........38 5.5.3 Adjust Parameters ........39 5.5.4 Applying the Specimen ........39 5.5.5 Blotting ............39 5.5.6 Freezing ............40 5.5.7 Transfer to Cryo Grid Box ......40 5.6 Transferring a Cryo-Grid Box.......41 5.7 Shutting Down and Bake Out ......41 5.8 Settings ............42 5.8.1 Tab “Process”...

  • Page 7: Introduction

    1.2 Field of Application and Intended Use any damage caused by the usage of such substances. This product, the EM GP2, an automatic plunge Finally, users must operate this product under freezer, is a dedicated cryo sample preparation the work area conditions outlined in this system for cryo electron microscopy.

  • Page 8: Symbols And Warnings

    1. Introduction technical but non-operational tasks (e.g. work NOTICE area set-up, transportation, decommissioning, and disposal) must be accordingly qualified. NOTICE indicates information considered Personnel interacting indirectly with this important but not hazard-related. product (e.g. laboratory personnel working in the proximity) and additional employees SAFETY without complete knowledge of the safety rules INSTRUCTIONS...

  • Page 9: General Safety Information

    (Section 7). always well ventilated when large quantities of nitrogen are evaporated (e.g. when transferring The EM GP2 was tested in accordance to IEC/ ). To dispose of residual LN , dump it into an EN 61010-1 and EN 61326-1. It complies with the...

  • Page 10: Safe Storage

    1. Introduction with higher evaporation rates present a safety DANGER risk and should be removed or subjected to maintenance. Regardless of their state, never Call Ambulance leave LN containers in closed storage rooms. Bring person to well ventilated area, call an ambulance and CAUTION apply artificial respiration! Evaporation...
  • Page 11 1. Introduction entering the aforementioned equipment does at hand temperature. In case of serious burns, not evaporate quickly and causes serious burns arrange for a skin specialist to see them at once. before there is time to remove it. In case of LN affecting the eyes, immediately rinse with water at hand temperature and Instead, always use protective glasses with side...

  • Page 12: Safety Measures Regarding Ethane And Propane

    1. Introduction colourless, odourless, and flammable. Exposure • Use only metal Dewar vessels designed can cause headache, nausea, vomiting, dizziness for LN and light-headedness. Similarly to LN , very high • Control the evaporation rate of the Dewar levels can cause death by asphyxiation from lack regularly of oxygen.

  • Page 13: Equipment And Handling

    1. Introduction 1.7.3 Equipment and Handling Physical Properties This product’s secondary cryogen container has • Understand the hazards of ethane and a volume of 2.5 ml. When working with ethane propane and propane, use adequate Personal Protective • Ensure rooms with ethane and propane Equipment (PPE).

  • Page 14: Product Description

    9 Secondary cryogen liquefier 10 Cryo transfer container 11 Foot switch (optional) 12 Touch panel The EM GP2 is a dedicated cryo-preparation system for plunge freezing cryo-fixation of thin samples on EM grids. The instrument was designed to make the preparation of vitreous frozen thin films straightforward and reproducible.

  • Page 15: Dimensions And Weight

    GN The EM GP2 is delivered in one box with the box for storage or transfer to the next instrument. following dimensions and weight: •...

  • Page 16: Stereomicroscope

    • Sec. cryogen capacity: 2.5 ml flow 50%, Dewar covered with lid, secondary cryogen temperature -180°C 2.4 Protective Measures To improve safety by reducing the occurrence and severity of residual hazards, the EM GP2 designed with constructive, technical, and informative protective measures. These measures are expected to function throughout the product’s lifetime without monitoring nor...

  • Page 17: Informative Protective Measures

    Conditions: Follow safety and shall be kept as such for further reference. instructions in Sections 1.6 and 1.7! The EM GP2 contains the following pictograms and safety signals. WARNING Suffocation Hazard Use in a well-ventilated area, even during power outage!

  • Page 18: Type Label

    2. Product Description 2.5 Type Label The type label is located on the rear side of the product, above the mains supply switch and cable connectors. See Section 9 for all technical specifications.

  • Page 19: Transportation And Storage

    3. Transportation and Storage 3.1 Transportation 3.3 Storage To prepare the EM GP2 for transport over large In cases when the device needs to be kept un- distances, empty the humidifier tank and the drip operational for a prolonged period, clean the tray (Section 5.4.3), and remove the secondary...

  • Page 20: Installation

    (± 10%) and that it is equipped with the required Under consideration of the safety measures safety measures and personal protective in Sections 1.6 and 1.7, install the EM GP2 in a equipment (Section 1.6). Installing an oxygen well-ventilated room of adequate volume. Make sensor in the room is highly recommended.

  • Page 21: Assembly

    The packing material is designed explicitly for the EM GP2 and shall be retained in case the instrument needs to be transported in the future. The product can sustain damage if it is not transported in the original packing.

  • Page 22: Accessories

    4.4 Accessories The main outfit of the EM GP2 consists of the basic instrument with a touch panel and an accessories kit as shown in the next figure. During the installation process, the authorized...

  • Page 23: Operation

    5. Operation 5. Operation 5.1 Switching on and User Interface Turn on the power with the main switch at the rear of the main unit. After the instrument boots up, the following screen will appear: After touching the screen, the instrument continues to initialise.

  • Page 24: Parameters

    5. Operation chamber (temperature, relative humidity and window heater) and the cryogen (temperature and GN flow). The Main Screen contains the following panels and functional keys: 1 The status panel displaying environment, filter paper, and cryogen parameters, alternating 5.2.1.1 Chamber Temperature periodically between actual and set values.

  • Page 25: Chamber Rel. Humidity

    5. Operation NOTICE Humidity The instrument will only actively increase humidity. Like for the temperature control the humidity regulation can be switched on or off with the button besides “Set”. When it is switched off, the warnings regarding the humidifier tank filling The chamber temperature regulation can be level (see below) will not be displayed either.

  • Page 26: Gn Flow

    5. Operation The forceps with the empty grid are always plunging and may vary from one gas supply to attached to the instrument in the “home” another, depending on its quality. position. The first checkbox (“rotate grid before specimen application”) rotates to a 5.2.1.5 GN Flow 180°...

  • Page 27: Blot Parameters

    5. Operation A delay between application of the specimen/ and keying in the desired blot time (0 to 200 s). pressing, the “Blot” button, and the actual filter This parameter is highly specimen specific – for paper blotting can be set with the “Delay time 3 to 5 µl aqueous solution, 0.5 to 3.0 s blotting are before blotting –...

  • Page 28: Single Blotting With Sensor

    5. Operation left/right arrows to move the filter paper to the correct position in small steps (0.2 mm). This is when the filter paper is touching all the grid and preferably the tips of the forceps too. The stereomicroscope can help with determining this position accurately.

  • Page 29: Multiple Blotting

    5. Operation The default size of the blotting window (search environmental chamber) or bending of the grid, distance) does not require adjustment under this point standardises the distance from the normal conditions. It should be noted that the filter paper to the grid, provided that the sample sensor scans through the search distance on volume and spreading of the liquid are kept both sides of the Center position Therefore, the...

  • Page 30: Plunge / Transfer Parameters

    The post-blotting time is desired by some operators to allow the water layer even out over In the above example, the EM GP2 will blot three the surface of the grid. It requires the humidifier times in the home position and three times in to be set up properly to avoid uncontrolled the 180°...

  • Page 31: Programs

    Entirely depending on the user’s personal instrument is switched off. preference, the EM GP2 allows to The program list can be accessed via the • Completely skip the transfer position (“Skip “Programs” button on the main screen.

  • Page 32: Editing Programs

    5. Operation 5.3.2 Editing Programs 5.3.4 Deleting Programs Programs can be edited by directly accessing To remove a program, go to the program list, the individual parameters from the main screen select the respective program and press as described in section 5.2. “Delete”.

  • Page 33: Humidifier

    5. Operation It should be noted that the automatic blotting assumed that a new filter paper will be set in at depends on the type of filter paper used. The that point (warning message at start up). sensor is set to recognize blotting of a drop of at Once all 10 positions on the filter paper are used least 3 µl by filter paper of the type Whatman ™...
  • Page 34 5. Operation When the filling of the humidifier tank falls below its minimum level during operation the system will beep and indicate that refilling is needed. When the humidifier tank is filled to its maximum, a warning will be displayed together with a warning beep asking to stop the filling.

  • Page 35: Emptying The Drip Tray

    Now pour liquid nitrogen (LN ), directly into the EM GP2 Dewar using a Dewar or similar. It will take around 1.8 litres LN to cool the Dewar and fill it to 100%.

  • Page 36: Filling The Secondary Cryogen

    Insert the gas liquefier in the chamber over the rises above -100 °C and starts to evaporate. secondary cryogen container while cooling When secondary cryogen is in the freezing down the EM GP2 with LN and wait until the chamber during refill, do not spill LN over the temperature of the cryogen container drops to or surface of the secondary cryogen –...

  • Page 37: Freezing A Grid

    The snap-lock forceps have a quick release Frostbite mechanism for rapid loading and easy unloading. Avoid contact with any residual The forceps fit onto the EM GP2 in only one cryogen liquid that might be still direction to maintain the orientation of the in the system.

  • Page 38: Lower Chamber

    EM GP2. The chamber will lower and if a 180° rotation for loading has been set up, the forceps will rotate by 180°.

  • Page 39: Adjust Parameters

    5. Operation For applying the specimen, a type of microliter When the environmental chamber is in the lower pipette must be chosen that can easily reach the position, the shutter closing the feedthrough to grid without getting caught at the ports. the cryogen will wiggle in regular intervals to prevent freezing to the bottom of the chamber.

  • Page 40: Freezing

    5.5.6 Freezing If “automatically plunge after blotting” is selected in the parameters (section 5.2.4, page 34), the EM GP2 will automatically plunge the grid into the secondary cryogen after blotting (and the elapse of the optional post-blot delay). If this option is not enabled, the user must press the “PLUNGE”...

  • Page 41: Transferring A Cryo-Grid Box

    To commence another run, press “Load Forceps” container completely with LN . Move the grid in the workflow and the environmental chamber box cryo transfer container back to the EM GP2 will rise for the next run. Dewar and resume freezing. 5.6 Transferring a Cryo-Grid Box 5.7 Shutting Down and Bake Out...

  • Page 42: Settings

    Dewar is completely empty. Therefore, the total On completion of the cryo work, the EM GP2 must bake out time will depend on the LN level and...

  • Page 43: Tab "Process

    5. Operation can be reduced to approx. 1/3. The check boxes 5.8.1 Tab “Process” in the lower right part of the “Process” tab allow to enable/disable this slow-down for the down and the up movements separately. 5.8.2 Tab “System” “Ext. remote alarm on” enables/disables a remote alarm signal via a jack plug on the rear of the main instrument.
  • Page 44 5. Operation them and send them to your service technician if asked to do so. The “Update” and “Service” buttons will be explained in the next section.

  • Page 45: Maintenance And Repairs

    6.1 Service Menu 2. Copy this file into the root directory of the USB-Stick The EM GP2 has a Service button under the 3. Extract the contents of the ZIP file into the root Settings menu (Section 5.8.2) which allows directory of the UBS-Stick (i.e.

  • Page 46: Cleaning And Upkeep

    To reduce contamination, it is recommended to use gloves when handling in the interior of the chamber! Before cleaning, switch off and unplug the EM GP2 from the mains power supply. WARNING If there is an available update, always update Disconnect Power “User Interface”...

  • Page 47: Cleaning The Humidifier Tank

    90ml of 70% ethanol, leave it for 15 minutes, drain, and rinse with distilled water. If the EM GP2 is left unused for longer periods of time, detach and empty the humidifier tank as described below and store it in a dry place.

  • Page 48: Disposal

    To prepare the EM GP2 for decommissioning, human pathogenic substances inside the device. empty the humidifier tank (Section 6.3.1) and Failure to do so might endanger the WEEE drip tray (Section 5.4.3), and remove any leftover recycling company’s personnel.

  • Page 49: Troubleshooting

    8.2 Warranty reference code on the first column– all possible warning and error messages displayed by the The EM GP2 is covered by a warranty according EM GP2, whereby to the sales conditions. If functional errors should • Warnings (coded WXXXX) refer to situations...
  • Page 50 The third column displays the warning or error’s description (sometimes including system status or short advice) as displayed by the software upon appearance. The fourth column provides additional information leading to the probable causes of the warning or error message. EM GP2 – Errors and Warnings Code Type / Decision...
  • Page 51 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information The chamber movement Shown after pressing “STOP” Warning was stopped. Press <OK> to during chamber movement. <OK> W1005 continue with the chamber <OK>/<Cancel> resumes movement.
  • Page 52 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information Confirmation request to delete Warning Are you sure you want to a program; <OK> proceeds with W1013 delete this program? deletion, <CANCEL> keeps the <OK>/<Cancel>...
  • Page 53 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information Shown when there’s no Dewar controller communication between the user Error E10E2 communication error. interface and the Dewar controller. <OK> Contact service! First, restart the system. If the issue persists, contact a service engineer.
  • Page 54 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information The blotter is in an undefined Blotter axis is not initialized. state; remove any obstacles in Error E2002 <OK> start initialization the chamber and press <OK> to <OK>/<Cancel>...
  • Page 55 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information Chamber axis initialization The chamber could not reach the Error failed. Could not reach upper top position; remove any obstacles E4001 end switch. <OK> start <OK>/<Cancel>...
  • Page 56 8. Troubleshooting EM GP2 – Errors and Warnings Code Type / Decision Message Further Information Plunger axis initialization The plunger retract did not Error failed. Could not reach upper reach the top position; remove E6001 end switch. <OK> start any obstacles and press <OK>...

  • Page 57: Technical Data

    9. Technical Data 9. Technical Data APPROVAL Conformity Marking CE (European Union) ELECTRICAL Mains Voltage 100/115/230 V (+/- 10%), 50–60 Hz Power Consumption ~ 220 W Control Voltage 24 V DC DIMENSIONS AND WEIGHT WIDTH x DEPTH x HEIGHT WEIGHT Basic Unit 336 x 445 x 883 mm ~ 45 kg...
  • Page 58 9. Technical Data DEWAR Dewar Volume Dewar Temperature -196 °C Dewar bake-out Temperature Max. +110°C LN2 Consumption ~ 1 l/h flow 50%, Dewar covered with lid, secondary cryogen temperature -180°C PREPARATION CHAMBER Allowed Secondary Cryogen Ethane / Propane Secondary Cryogen Container 0 to -196 °C Temperature Secondary Cryogen Container...
  • Page 59 To download the EC Declaration of Conformity, use this link https://www.leica-microsystems.com/products/sample-preparation-for-electron-microscopy/p/leica- em-gp2/downloads/ If you have any further questions or require technical support, please contact your country’s Leica branch office or your local contact person directly. The appropriate contacts can be found on the Internet under:...

  • Page 60: Connect With Us

    CONNECT WITH US! Leica Microsystems CMS GmbH | Ernst-Leitz-Strasse 17–37 | D-35578 Wetzlar (Germany) Tel. +49 (0) 6441 29-0 | F +49 (0) 6441 29-2599 www.leica-microsystems.com...

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