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Related Manuals for Thermo Scientific Invitrogen Countess II FL
Summary of Contents for Thermo Scientific Invitrogen Countess II FL
- Page 1 ™ Countess II FL automated cell counter USER GUIDE For fluorescence and brightfield applications Catalog Number AMQAF1000 Document Part Number MP10826 Publication Number MAN0010644 Revision E.0 For Research Use Only. Not for use in diagnostic procedures.
- Page 2 Life Technologies Corporation | 22025 20th Ave SE Ste. 100 | Bothell, WA 98021 For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition. The information in this guide is subject to change without notice. DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.
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Page 3: Table Of Contents
Contents ■ CHAPTER 1 About this guide ..........7 Audience . - Page 4 Contents ■ CHAPTER 4 Cell count and cell viability assays ..... . 23 Count cells in brightfield ............23 Capture and count .
- Page 5 Contents ■ CHAPTER 7 Save results ..........44 Save count results .
- Page 6 Contents ■ APPENDIX C Ordering information ........66 ™...
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Page 7: Chapter 1 About This Guide
About this guide Audience This user guide is for laboratory staff operating, maintaining, and analyzing data using the Countess ™ II FL Automated Cell Counter. User documentation ™ The guides listed below are available for the Countess II FL Automated Cell Counter. -
Page 8: User Attention Words
Chapter 1 About this guide User attention words User attention words Two user attention words appear this document. Each word implies a particular level of observation or action as described below. Note: Provides information that may be of interest or help but is not critical to the use of the product. -
Page 9: Chapter 2 Product Information
Product information Product contents ™ The Countess II FL Automated Cell Counter is shipped with the following components. Component Quantity Countess ™ II FL Automated Cell Counter (Cat. No. AMQAF1000) 1 each Power Cord with 4 adaptor cords 1 each (for U.S./Canada/Taiwan/Japan, Europe, or UK) Countess ™... -
Page 10: Product Description
Chapter 2 Product information Product description Product description Countess II FL ™ ™ The Countess II FL Automated Cell Counter is a fully automated, 3-channel cell counter and assay platform that uses EVOS ™ light cube technology, state-of-the-art automated cell optics, and image analysis algorithms to analyze fluorescently labeled cells or trypan counter blue stained samples in suspension. -
Page 11: Instrument Exterior Components
Chapter 2 Product information Instrument exterior components Instrument exterior components Touch-screen display: The 7‑inch capacitive touch-screen display is the main user interface of the Countess II FL Automated Cell Counter. It contains the buttons for all ™ instrument functions and displays data from the cell count. USB ports: The USB ports allow you to transfer and save the cell count data and image to an external computer for record keeping and printing purposes. -
Page 12: Chapter 3 Getting Started
Getting started Installation Operating • Place the instrument on a level surface away from vibrations emanating from other pieces of equipment. environment • Allow at least 5 cm (2 in) free space at the back of the instrument to allow for proper ventilation and prevent overheating of electronic components. -
Page 13: Turn On The Instrument
Chapter 3 Getting started Installation Turn ON the 1. Turn on the instrument by flipping the power switch on the back of the instrument (“Instrument exterior components“ on page 11) to the ON position. instrument The instrument initializes and displays the Home screen. 2. -
Page 14: Load Profile
Chapter 3 Getting started Load profile Load profile Profiles screen Profiles screen allows you to create and save up to 9 customized profiles. Each custom profile defines the count parameters (size, brightness, circularity, and fluorescence intensity) and automatic instrument functions (Auto Lighting and Auto FL Threshold) for a consistent and streamlined workflow. -
Page 15: Count Parameters
Chapter 3 Getting started Load profile Count parameters Count parameters are adjusted in the Edit profile screen using the parameter sliders. Parameter sliders correspond to a single channel, which is selected using the channel selection radio buttons located above the sliders. •... -
Page 16: Load A Profile
Chapter 3 Getting started Load profile Load a profile 1. Press the Profiles button located on the upper left corner of the screen to open the Profiles screen. 2. Press the desired profile to select, and then press Load. The instrument will load the count parameters specified in the selected profile and return to the previous screen. - Page 17 Chapter 3 Getting started Load profile Define count parameters for the fluorescent channels 1. To define the new count parameters for fluorescence assays (available only for ™ instruments equipped with the optional EVOS light cubes): a. From the FL selection box, select the desired channel using the corresponding radio button.
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Page 18: Display Of Profile Names
Chapter 3 Getting started Load profile 2. To assign a name to the new profile or to change the name of the existing profile, press the Profile name text box. The alpha-numeric keypad opens. 3. Type in the desired profile name using the alpha-numeric keypad. To enter symbols, press the symbol (@%&) key. -
Page 19: Prepare Sample
Chapter 3 Getting started Prepare sample Prepare sample Recommendations To obtain the best results, follow these recommendations: • Ensure that the cell sample is homogeneously mixed. • The measurement range extends from 1 × 10 –1 × 10 cells/mL, but the optimal range is 1 ×... -
Page 20: Load Countess ™ Ii Fl Reusable Slide
Chapter 3 Getting started Slide operation Load Countess ™ 1. Before loading your sample into the Countess ™ II FL Reusable Slide, place a cover slip on the counting chamber, making sure the cover slip is clean and free FL reusable slide of grease. -
Page 21: Countess ™ Ii Fl Reusable Slide
Chapter 3 Getting started Slide operation 2. Load the chamber slide with your sample as described in “Load Countess ™ chamber slide“ on page 19, and then insert the slide into the slide carrier in the slide port until it clicks into place. 3. - Page 22 Chapter 3 Getting started Slide operation 3. Load the reusable glass slide with the sample as described in “Load Countess ™ FL reusable slide“ on page 20, and place the loaded slide into the white slide carrier. 4. Insert the carrier and reusable slide assembly into the slide port, and gently push into the instrument until it clicks into place.
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Page 23: Chapter 4 Cell Count And Cell Viability Assays
Cell count and cell viability assays Count cells in brightfield Capture and count 1. Prepare the sample by adding 10 µL of your cell suspension to 10 µL of 0.4% trypan blue stain. Mix the sample mixture well by pipetting up and down a few times. - Page 24 Chapter 4 Cell count and cell viability assays Count cells in brightfield 7. Press the Set button to set the focus and collapse the focus controls. Once the focus has been set, the Set button on the focus slider becomes inactive, confirming that the focus setting has been stored.
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Page 25: Next Steps
Chapter 4 Cell count and cell viability assays View results Next steps • To identify the objects (i.e., cells) counted as Live or Dead, press More to go to the ™ Advanced screen (“Identify objects counted“ on page 26). • To see the distribution of live and dead cells in a graphical format, press the Graph button (“Graph count results“... -
Page 26: Identify Objects Counted
Chapter 4 Cell count and cell viability assays Identify objects counted Identify objects counted Advanced screen ™ ™ The Advanced screen allows you to identify the objects (i.e., cells) counted in each channel and included in the count results for further review. After reviewing the marked objects, you can adjust the threshold for size, brightness, and/or circularity as desired for your application (“Gate count results“... -
Page 27: Graph Count Results
Chapter 4 Cell count and cell viability assays Graph count results Graph count results View graph For cell count and cell viability assays performed in the brightfield channel, you can view the distribution of cells (live and/or dead) based on size in a graphical format. ™... - Page 28 Chapter 4 Cell count and cell viability assays Gate count results 3. On the Advanced ™ screen, press Adjust to open the Adjust screen. 4. Optional: Press the Graph button to view the distribution of cells (live and/or dead) based on size as you gate the count results (“Save as new protocol“ on page 29).
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Page 29: Save As New Protocol
Chapter 4 Cell count and cell viability assays Save as new protocol Save as new protocol Edit and save 1. If you have made any changes to the count parameters before or after performing a count, the displayed profile name is appended with the (*) symbol and the profile as new Advanced ™... - Page 30 Chapter 4 Cell count and cell viability assays Save as new protocol 4. The Edit profile screen opens and displays the edited count parameters from the Adjust screen (“Gate count results“ on page 27). Note: If you have selected a profile that had been previously saved, the name of that profile populates the Profile name text box by default.
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Page 31: Chapter 5 Fluorescence Assays
Fluorescence assays Count cell fluorescence Overview Countess ™ II FL Automated Cell Counter equipped with the optional EVOS ™ light cubes can be used for a variety of fluorescent applications, including simultaneous counts of cells stained with two different fluorescent dyes, GFP and RFP expression, and apoptosis and cell viability assays. - Page 32 Chapter 5 Fluorescence assays Count cell fluorescence 6. Optional: To manually adjust the focus, press the Focus ™ button and use the Focus ™ slider to bring your sample into focus as described in “Set nominal focus“ on page 57. 7.
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Page 33: Next Steps
Chapter 5 Fluorescence assays Count cell fluorescence 10. Press the light source button for the channel you wish to set exposure and adjust the exposure using the light source slider. Repeat the procedure for the remaining channels, if desired. 11. After setting the exposure, press Done to return to the Capture screen. To return to the Capture screen without changing the exposure, press Cancel. -
Page 34: View Results
Chapter 5 Fluorescence assays View results • To permanently save the results, press Save (Chapter 7, “Save results“). • To perform a new count, remove the slide and reinsert it second chamber first into the instrument, or insert a new sample slide. View results Results screen for The Results screen for cell fluorescence assays displays a composite image of the... -
Page 35: Identify Cells Counted In Fluorescence Assays
Chapter 5 Fluorescence assays Identify objects counted Identify cells 1. On the Results screen, click More to open the Advanced ™ screen. counted in fluorescence assays 2. Optional: To view your sample under a specific light source (brightfield and/or fluorescent), select the desired Channels checkbox (brightfield in the example above). -
Page 36: Graph Count Results
Chapter 5 Fluorescence assays Graph count results Graph count results View graph for cell For fluorescence assays, you have the option of viewing the distribution of the cells based on size or based on relative fluorescence intensity in a graphical format. fluorescence assays ™... -
Page 37: Gate Count Results
Chapter 5 Fluorescence assays Gate count results 3. To remove the cells counted in a specific channel from the graph, uncheck the corresponding channel check box on the graph. The graph automatically updates and displays the distribution of cells based on relative fluorescence intensity only in the selected (i.e., checked) channel. - Page 38 Chapter 5 Fluorescence assays Gate count results 4. Press Adjust to open the Adjust count parameters screen, which contains the controls for adjusting the count parameters in the selected channel. Note: For a description of the count parameters and count parameter controls (i.e., parameter sliders), see “Count parameters“...
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Page 39: Save As New Protocol
Chapter 5 Fluorescence assays Save as new protocol Save as new protocol Edit and save 1. If you have made any changes to the count parameters, the displayed profile name is appended with the (*) symbol and the Advanced ™ results screen profile as new displays the New protocol button, which allows you to save the changes to the... - Page 40 Chapter 5 Fluorescence assays Save as new protocol 4. The Edit profile screen opens and displays the edited count parameters from the Adjust screen (“Gate count results“ on page 37). Note: If you have selected a profile that had been previously saved, the name of that profile populates the Profile name text box by default.
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Page 41: Chapter 6 Dilution Calculator
Dilution calculator Calculate dilution Dilution calculator Dilution calculator function allows you to calculate the volume of cell sample and buffer needed to reach a desired concentration using the count results. 1. On the Results screen, press Dilution Calculator to open the Dilution calculator Calculate dilution screen. - Page 42 Chapter 6 Dilution calculator Calculate dilution • For fluorescence assays: Channel 1, Channel 2, or Total cells (Channel 1 + Channel 2). By default, Total cells is selected. Note: Channel 1 and Channel 2 name displayed in the dropdown menu ™...
- Page 43 Chapter 6 Dilution calculator Calculate dilution 4. Press the total volume text box (How many mL do you need?), then enter the total volume of the sample you wish to have at the new concentration using the number pad. By default, the total volume box is blank. The maximum volume you can enter is 999.9 mL and you can use only a single decimal.
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Page 44: Chapter 7 Save Results
Save results Save count results Save screen The Countess ™ II FL Automated Cell Counter allows you to save your data and images using a USB flash drive. To save your experiment, choose from the following options, in any combination: •... - Page 45 Chapter 7 Save results Save count results 2. On the Results screen, press Save to go to the Save screen. 3. To assign a name to your experiment, press the File name text field. The alpha- numeric keypad opens. 4. Enter the file name using the alpha-numeric keypad. To enter symbols, press the symbol (@%&) key.
- Page 46 Chapter 7 Save results Save count results 6. Select the desired mode to save your experiment (Result, Images, Data, Report). You can select an individual mode (e.g., Result only) or any combination of modes (e.g., Result, Images, Data, and/or Report). In the example below, Data and Report are selected.
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Page 47: Report
Chapter 7 Save results Report 9. Press Save to save your experiment in the selected mode(s) in the USB drive. 10. Press Close and then transfer the USB drive to the desired location. Report Report file The Report function allows you to save a printer-friendly report of the results, graphs, and images in the selected format (PDF, PNG, or JPEG). - Page 48 Chapter 7 Save results Report Report from brightfield count • The top section of the Report contains a table with the results as displayed on the Countess ™ Results screen, showing the concentration of the sample, and the percentage and number for the total, Live, and Dead channels. •...
- Page 49 Chapter 7 Save results Report Report from fluorescence count • The top section of the Report contains a table with the results as displayed on the Countess Results screen, showing the concentration of the sample, and the percentage ™ and number of cells for the total, FL1, FL2, and FL1 + FL2 channels. •...
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Page 50: Chapter 8 Instrument Settings
Instrument settings Overview Instrument To access the Instrument Settings screen, press the Instrument Settings button on the Home page (“Turn ON the instrument“ on page 13). settings screen In the Instrument Settings screen, you can: • perform software update (“Software update“ on page 50) •... -
Page 51: Update The Countess ™ Ii/Ii Fl Software
Chapter 8 Instrument settings Software update Update the 1. Go to www.thermofisher.com/countessupdate, and download the latest ™ Countess II/II FL software version to your desktop. Countess ™ II/II FL software Note: The software update file has a version-specific name followed by the ™... -
Page 52: Date/Time
Chapter 8 Instrument settings Date/Time Date/Time Set the date and 1. Press the Instrument Settings button on the Home page (“Count parameters“ on page 15) to open the Instrument Settings screen. time 2. Press Date/Time on the Instrument Settings menu to open the Date/Time screen. 3. - Page 53 Chapter 8 Instrument settings Date/Time 7. Select the Time format you wish to use. Available options are 12 Hour and 24 Hour formats. 8. Press any Time text box (Hours or Minutes) to open the Edit Time keypad. 9. Using the keypad, enter the time into Hours and Minutes text boxes, pressing Enter after each entry.
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Page 54: Change Light Cube
Chapter 8 Instrument settings Change light cube Change light cube ™ ™ Install or change The Countess II FL Automated Cell Counter can accommodate up to two EVOS ™ light cubes. Each user-interchangable, auto-configured EVOS light cube contains an EVOS ™... - Page 55 Chapter 8 Instrument settings Change light cube 7. Using the tool provided on the inside of the back panel (Figure A), secure the light cube by tightening the two screws on the end of the cube (red arrows in Figure B). 8.
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Page 56: Chapter 9 Maintenance
Maintenance Instrument care General • Use the appropriate cleaning solutions for each component, as indicated in the cleaning procedures in “Clean the Countess II FL automated cell counter“ on ™ guidelines for care page 56. • If liquid spills on the instrument, turn off the power immediately and wipe dry. Power supply Always use the correct power supply. -
Page 57: Clean The Touch-Screen
Chapter 9 Maintenance Set nominal focus Clean the touch- • Wipe the touch-screen of the Countess ™ II FL Automated Cell Counter using a soft, lint-free cloth moistened with an LCD cleaning solution. Do not apply screen excessive force during cleaning. Wipe the touch-screen dry immediately after cleaning. -
Page 58: Set Nominal Focus
Chapter 9 Maintenance Set nominal focus To enable optimal auto focus functionality, you may need to initially refine the brightfield focus by adjusting it manually and then setting the nominal focus. This allows the auto focus function to have a set point from which to focus on the cells in subsequent samples. - Page 59 Chapter 9 Maintenance Set nominal focus 6. Use the Focus ™ slider or the plus and minus buttons to refine the brightfield focus. Note: If needed, Zoom in on the image to adjust focus or lighting. 7. After finding the optimal focus, press Set to set the nominal focus. Once the focus has been set, the Set button on the focus slider becomes inactive, confirming that the focus setting has been stored.
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Page 60: Appendix A Troubleshooting
Troubleshooting Note: The software for the Countess ™ II FL is updated regularly. If you are having any issues with your experiments, first check the website to see if a new software version is available. You can download the most recent version of the software from ™... - Page 61 Appendix A Troubleshooting Set nominal focus Problem Possible solutions • The beads can settle quickly in solution, which will ™ Getting incorrect concentration for the Countess test affect the concentration reading. beads • Vortex the bead stock on high for a full 30 seconds to resuspend, and add 10 µL of the bead suspension to 10 µL of trypan blue without delay.
- Page 62 Appendix A Troubleshooting Set nominal focus Problem Possible solutions • Ensure that the cells are focused correctly so that Abnormally high percentage of dead cells or live cells live cells have bright centers and dead cells are dark counted as dead throughout (see “Set nominal focus“...
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Page 63: Appendix B Product Specifications
Product specifications Technical specifications Physical Instrument type: Benchtop cell counter and suspension cell- characteristics based assay platform Instrument dimensions: 9 (W) × 5½ (D) × 9 inches (H) Weight: 8 lbs Operating power: 100–240 VAC, 0.58 A MAX Frequency: 50/60 Hz Electrical input: 12 VDC, 2 A Installation site:... -
Page 64: Evos ™ Light Cubes
Appendix B Product specifications EVOS light cubes ™ ™ EVOS light cubes LED illumination ™ The Countess II FL Automated Cell Counter utilizes an adjustable intensity LED light source provided by the proprietary, user-interchangeable LED light cube (see “EVOS light cubes“ on page 64). Because the LED light source is as close as possible ™... - Page 65 Appendix B Product specifications EVOS light cubes ™ Note: The EVOS ™ light cubes are available only for the Countess ™ II FL Automated ™ Cell Counter. The Countess II Automated Cell Counter uses only brightfield ™ illumination and does not support the EVOS light cubes.
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Page 66: Appendix C Ordering Information
Ordering information ™ Countess II FL automated cell counter and accessories ™ The following Countess II FL instruments and instrument accessories are available from Thermo Fisher Scientific. For more information, visit www.thermofisher.com or contact Technical Support. Product Quantity Cat. No. Countess ™... - Page 67 Appendix C Ordering information Accessory products Product Quantity Cat. No. ™ Countess Test Beads (1 × 10 beads/mL) 1 mL C10284 Trypan blue stain (0.4 %) 2 × 1 mL T10282 Each box of Countess ™ Cell Counting Chamber Slides contains 50 slides and 2 × 1 mL vials of trypan blue (0.4%), sufficient for 100 counts.
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Page 68: Appendix D Csv File Format
CSV file format CSV file format, explained Overview A comma-separated values (CSV) file stores tabular data (numbers and text) in plain- text form. Plain text means that the file is a sequence of characters, with no data that has to be interpreted as binary numbers. A CSV file can be opened with any third party software or spreadsheet program. - Page 69 Appendix D CSV file format CSV file format, explained Category Column Name Description Fluorescence Cube 2 concentration Concentration of cells showing fluorescence in the second cube position Cube 2 (%) Percentage of the total cells in brightfield that show fluorescence in the second cube position Cube 2 cells counted Total number of cells counted in the second cube position...
- Page 70 Appendix D CSV file format CSV file format, explained Category Column Name Description Fluorescence Count BF Brightness min “Brightfield” adjustment slider value for minimum Parameters brightness BF Brightness max “Brightfield” adjustment slider value for maximum brightness BF Circularity “Brightfield” adjustment slider value for circularity Cube 1 Brightness min First (top) light cube adjustment slider value for minimum brightness...
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Page 71: Appendix E Safety
Safety WARNING! GENERAL SAFETY. Using this product in a manner not specified in the user documentation may result in personal injury or damage to the instrument or device. Ensure that anyone using this product has received instructions in general safety practices for laboratories and the safety information provided in this document. -
Page 72: Electrical Symbols
Appendix E Safety Electrical symbols Electrical symbols The following table describes the electrical symbols that may be displayed. Symbol Description Indicates the On position of the main power switch. Indicates the Off position of the main power switch. Indicates a standby switch by which the instrument is switched on to the Standby condition. -
Page 73: Environmental Symbols
Appendix E Safety Environmental symbols Symbol Description Caution, hot surface or other high-temperature hazard. Caution, laser. Caution, moving parts. Caution, potential biohazard. Caution, ultraviolet light. Environmental symbols The following symbol applies to all Thermo Fisher Scientific electrical and electronic products placed on the European market after August 13, 2005. Symbol Description Do not dispose of this product as unsorted municipal waste. -
Page 74: Safety Labels On Instruments
Appendix E Safety Safety labels on instruments Safety labels on instruments The following CAUTION, WARNING, and DANGER statements may be displayed Thermo Fisher Scientific instruments in combination with the safety symbols described in the preceding section. Symbol English Français CAUTION! Hazardous chemicals. Read the Safety ATTENTION! Produits chimiques dangereux. -
Page 75: Safety Precautions
Appendix E Safety General instrument safety Safety precautions • Do not install the instrument in heavy humidity, such as a greenhouse or an incubator, to avoid a danger of electric shock. If water or other material enters the instrument, the adaptor, or power inlet, disconnect the power cord and contact a service person. -
Page 76: Chemical Safety
Appendix E Safety Chemical safety Chemical safety WARNING! GENERAL CHEMICAL HANDLING. To minimize hazards, ensure laboratory personnel read and practice the general safety guidelines for chemical usage, storage, and waste provided below. Consult the relevant SDS for specific precautions and instructions: ·... -
Page 77: Chemical Waste Safety Guidelines
Appendix E Safety Electrical safety Chemical waste To minimize the hazards of chemical waste: safety guidelines • Read and understand the Safety Data Sheets (SDSs) provided by the manufacturers of the chemicals in the waste container before you store, handle, or dispose of chemical waste. -
Page 78: Biological Hazard Safety
Appendix E Safety Biological hazard safety Part Symbol Description Power DANGER! ELECTRICAL HAZARD. Grounding circuit continuity is vital for the safe operation of equipment. Never operate equipment with the grounding conductor disconnected. DANGER! ELECTRICAL HAZARD. Use properly configured and approved line cords for the voltage supply in your facility. DANGER! ELECTRICAL HAZARD. -
Page 79: Safety And Electromagnetic Compatibility (Emc) Standards
Appendix E Safety Safety and electromagnetic compatibility (EMC) standards In the EU: • Check your local guidelines and legislation on biohazard and biosafety precaution, and the best practices published in the World Health Organisation (WHO) Laboratory Biosafety Manual, third edition www.who.int/csr/resources/publications/biosafety/WHO_CDS_CSR_LYO_2004 _11/en/ Safety and electromagnetic compatibility (EMC) standards... -
Page 80: Documentation And Support
Documentation and support Customer and technical support Visit thermofisher.com/support for the latest service and support information. • Worldwide contact telephone numbers • Product support information – Product FAQs – Software, patches, and updates – Training for many applications and instruments •... - Page 82 thermofisher.com/support | thermofisher.com/askaquestion thermofisher.com 7 June 2019...
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