Fluidigm Polaris User Manual
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Fluidigm Polaris User Manual

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PN 100-9580 E1
Polaris
USER GUIDE

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  • Page 1 PN 100-9580 E1 Polaris USER GUIDE...
  • Page 2 © 2018 Fluidigm Corporation. All rights reserved. 6/2018 For technical support visit fluidigm.com/support. North America +1 650 266 6100 | Toll-free (US/CAN): 866 358 4354 | techsupport@fluidigm.com Latin America +1 650 266 6100 | techsupportlatam@fluidigm.com Europe/Middle East/Africa/Russia +44 1223 598100 | techsupporteurope@fluidigm.com Japan +81 3 3662 2150 | techsupportjapan@fluidigm.com...

  • Page 3: Table Of Contents

    Contents Purpose Skip, Pause, Stop, or Abort a Step on Polaris Safety Alert Conventions Export Results to a USB Drive Export a Debug Log to a USB Drive Chapter 1: Introducing Polaris View the Results Exported to the USB Drive...

  • Page 4: Purpose

    Purpose This guide describes how to use the Polaris™ System, including the instrument and system software. For complete instrument specifications, see the Polaris Site Requirements User Guide (PN 100-9581). Safety Alert Conventions...
  • Page 5 Read and understand the SDSs before handling chemicals. To obtain SDSs for chemicals ordered from Fluidigm, either alone or as part of this system, go to fluidigm.com/sds and search for the SDS using either the product name or the part number.

  • Page 6: Chapter 1: Introducing Polaris

    You can see an example of the Results step for a Dose Response experiment in Run: View the Results on page 35. The following table is an example of fluorophores that can be used on Polaris: Table 1. Examples of cell-staining dyes...

  • Page 7: Components Of The Polaris System

    ON/OFF switch & power socket Display port (left) & Ethernet port (right) USB ports Premixed gas inlet Figure 2. Back of the Polaris instrument. There are also two USB 2.0 ports on the side of the instrument. Polaris: User Guide...
  • Page 8 Chapter 1: Introducing Polaris Components of the Polaris System Status indicator Environmental Control interface plate (EC IP) USB port (right side) Tray Figure 3. Open tray, exposing the EC IP above the IFC The Environmental Control Interface Plate (EC IP) resides on top of the IFC and provides an environment that is suitable for cell culture using user-desired gas composition.
  • Page 9 For more information about the reagents and cells for this IFC, see the Polaris mRNA Seq Protocol (PN 101-0082). Figure 6. The Polaris Single-Cell Dosing mRNA Seq IFC Notched corner Figure 7.

  • Page 10: Components Included In Shipping Box

    1/4 inch tubing, 50 feet Connects your CDA system to the compressed air inlet on the back of the Polaris instrument and connects the gas tank to the premixed gas inlet on the back of the Polaris instrument. NOTE To order additional items, contact Fluidigm sales.

  • Page 11: System Functions

    Regulatory Compliance The following directives and harmonized standards were used to evaluate the safety and performance of the Polaris system: General Regulations and Requirements • 2014/35/EU European Parliament Low Voltage Directive •...

  • Page 12: Conformity Symbols On The Instrument

    Indicates conformity with European Union requirements for safety and electromagnetic compatibility. Indicates conformity with North American requirements for safety and electromagnetic compatibility. Refer to the Polaris System Site Requirements Guide (PN 100-9581) for more detailed information on the recommended environmental conditions. Polaris: User Guide...

  • Page 13: Chapter 2: Getting Started With Polaris

    Chapter 2: Getting Started with Polaris This chapter shows you how to start and set up the Polaris™ instrument to image, select, capture, and dose or culture single cells individually. Start the Polaris System Plug the power cable into the power socket. Connect to the network.
  • Page 14 Chapter 2: Getting Started with Polaris Start the Polaris System The first screen that you see displays the initialization screen for roughly 10 to 15 seconds while all the system components are initializing: When initialization is complete, you see the next screen, which allows you to select...

  • Page 15: The Polaris Touchscreen

    Chapter 2: Getting Started with Polaris The Polaris Touchscreen The Polaris Touchscreen The LCD display is an interactive touchscreen that allows you to design experiments quickly, set up the instrument for use, and monitor progress during the current run. To access areas that are outside the current area on display, use your fingers and swipe to the right or left.

  • Page 16: The Polaris Menu

    Chapter 2: Getting Started with Polaris The Polaris Menu The Polaris Menu The Polaris menu is accessible before you begin the Prime step and after you exit the Results step: This menu allows you to create new experiments; view, export, or clone experiments;...

  • Page 17: Begin With A New Experiment

    For more information on how to run an mRNA Seq experiment, see the section Running a Single-Cell mRNA Seq Experiment on Polaris in the Polaris mRNA Seq Protocol (PN 101-0082). Begin with a New Experiment Tap the Fluidigm logo at the bottom-left of the touchscreen: When you see the Polaris menu on the left panel, tap New Experiment.

  • Page 18: Begin With An Existing Experiment

    Begin with an Existing Experiment Tap the Fluidigm logo at the bottom-left of the touchscreen: When you see the Polaris menu on the left panel, tap View Runs. On the Select a run to view screen, you can see your previous experimental setups and export completed run data (including capture annotation, intensities, and images).

  • Page 19: Set Up The Overview

    The lower area of the screen appears that allows you to make the change. Tap Suspension Cell Priming in the Priming Type box to change the value to Adherent Cell Priming: This action also changes the display of the Prime step to Adherent Cell Priming: Polaris: User Guide...

  • Page 20: Set Up The Hemocytometer Step

    Place the hemocytometer into the hemocytometer holder and place the holder into the Polaris tray with the notch facing toward you on the front-left side, and then place the EC IP over the top of the hemocytometer (EC IP not shown here):...
  • Page 21 After you tap Image on the Select parameters to focus and image screen, you see the Hemocytometer images acquired screen with the left and right sides of the hemocytometer on display for the selected Yellow channel. For example: Polaris: User Guide...
  • Page 22 (red) exhibit a value of fluorescence intensity of 12,000 and that all the cells on that side are within a range of fluorescence intensity between 7,000 and 31,000. Switching channels. Tap the channel of your choice located in the upper-left area of the screen. For example: Polaris: User Guide...
  • Page 23 Refocus. (Optional) View Hemocytometer Data Outside the Run If you had inserted at least a 3 GB USB key into the USB portal on Polaris before performing the Hemocytometer step, you can view the hemocytometer data by opening the MS Excel HemocytometerData.xlsx file on the USB drive.

  • Page 24: Set Up The Cell Selection Step

    The minimum cell input number is 12 cells/μL. Number of Cell Enter the number of cell populations to be selected Populations for the Dose and Feed or the No Treatment experiments. NOTE Not available in a Time Course experiment. Polaris: User Guide...

  • Page 25: Set Up The Cell Population Parameters

    IFC. See the recommended amount of input cells based on cell population purity in the Polaris mRNA Seq Protocol (PN 101-0082). Set Up the Cell Population Parameters...

  • Page 26: Set Up Cell Selection Parameters For Each Channel

    IFC must be re-imaged for the new exposure value to be applied. Tap Image. oad a new set of cells Tip: If needed, you can l into the IFC and image them. Tap Load. Polaris: User Guide...

  • Page 27: Set Up Cell Visibility Parameters For Each Channel

    Treatment groups 8 groups of 6 cells 6 groups of 8 cells Dosing delivery Simultaneous, from 48 inlets in Zone 2 Staggered, from inlets 6 through 13 Medium delivery Simultaneous, from R1 (upper-left From R1 (upper-left reservoir) reservoir) Polaris: User Guide...
  • Page 28 Blood Gas Disabled. Conditions A (Optional) Rename the assigned default name of through G each dosing group. Tap the Dose and Feed image. Tap anywhere in the dosing and feeding image beneath Dose and Feed: Polaris: User Guide...
  • Page 29 The lower area of the screen appears that allows you to make changes to the default parameters, as follows: Tap any box and change the default parameters if needed as described in Table When you are finished, tap anywhere outside the area of the dosing and feeding parameters area. Polaris: User Guide...
  • Page 30 Tap the Time Course image. Tap anywhere in the time course image beneath Time Course: The lower area of the screen appears that allows you to make changes to the default parameters, as follows: Polaris: User Guide...

  • Page 31: Set Up The Post Stain/Wash Step

    Select the number of channels to use for imaging. If you select the 0 Channel option, there will be no imaging. Channel Select the desired channels to be used for Yellow, Green, Far Red, Zombie Yellow post-stain imaging. Polaris: User Guide...

  • Page 32: Shut Down The Polaris System

    Shut Down the Polaris System Press and hold the power button on the front of the Polaris instrument to turn it on or off. If you are shutting down, this prompts the system to display the System Shutdown menu. Follow the instructions on the Polaris touchscreen to shut down the instrument properly.

  • Page 33: Unloading The Ifc And Ec Ip From Polaris

    Tap Start or Initialize. Unloading the IFC and EC IP from Polaris When it is time to remove the IFC from the Polaris system, Open appears. Tap Open. Remove the EC IP from the top of the IFC. Remove the IFC from the tray.

  • Page 34: Skip, Pause, Stop, Or Abort A Step On Polaris

    Setting Up the Experimental Run Skip, Pause, Stop, or Abort a Step on Polaris Skip, Pause, Stop, or Abort a Step on Polaris Available Actions Description Skip When you are allowed to skip to the next step to avoid the current step entirely, Skip appears.

  • Page 35: Export A Debug Log To A Usb Drive

    Export a Debug Log to a USB Drive This procedure exports all the data for an existing run into a *.zip file with subfolders where relevant to the USB drive that is connected to your Polaris instrument. This procedure assumes that you: •...

  • Page 36: View The Results Exported To The Usb Drive

    This folder contains images taken one time per hour in each channel during dosing. CellImages: HemocytometerImages This folder contains the MS Excel HemocytometerData.xlsx file with data and images for the fluorescent cells that were detected in the 2-sided hemocytometer. Polaris: User Guide...
  • Page 37 Stain/Wash step (the Stain & Wash option, but not the Wash Only option) of the experiment for each selected channel. CellImages: SelectionImages This folder contains a single image of all capture sites in each channel selected and a video file (.gif) of the selection process showing each selection event in sequence. Polaris: User Guide...
  • Page 38 Setting Up the Experimental Run View the Results Exported to the USB Drive Run.xlsx The Run.xlsx file contains a formatted summary of the experiment in MS Excel format. For example: Polaris: User Guide...
  • Page 39 Setting Up the Experimental Run View the Results Exported to the USB Drive Run.log The Run.log contains a log of all the events of the experiment in chronological order. For example: Polaris: User Guide...
  • Page 40 Setting Up the Experimental Run View the Results Exported to the USB Drive Run.tsv The Run file contains all the data for the run and can be opened as a tab-delimited file in Microsoft Excel. For example: Polaris: User Guide...

  • Page 41: Appendix A: Troubleshooting

    ON position. No display after power is Polaris system power Turn OFF the Polaris system power switch on the back turned on supply not fully reset panel. Wait 10 seconds and reboot by turning ON the power switch.

  • Page 42: Running

    “THIS SIDE UP” facing up. • Interface plate Confirm that the black vinyl film was removed from the missing bottom of the IFC. Technical • If the error persists, contact Fluidigm Support Unrecoverable error Various • Restart the system. Technical •...
  • Page 43 1 If steps 1–3 do not correct the problem, reboot by switching power OFF at the rear of the instrument, wait 10 seconds, switch power ON, and restart the run. (See Components of the Polaris System on page 7.) 2 If the system fails to recover, contact Fluidigm Technical Support...

  • Page 44: Appendix B: Maintenance, Decontamination, And Disposal

    Appendix B: Maintenance, Decontamination, and Disposal Cleaning and Maintenance This section describes how to clean and maintain your Polaris system for optimal performance. Before using a cleaning or decontamination method other than those recommended by Fluidigm, verify with Technical Support...

  • Page 45: Clean The Environmental Control Interface Plate

    We recommend cleaning the EC IP before use and before you store it in the Interface Plate Storage Container. If you need to clean the EC IP while it is in use, tap Open when you see that option on the touchscreen, and remove the interface plate from the tray. Polaris: User Guide...

  • Page 46: Clean The Touchscreen

    Decontamination and Disposal Ensure that the Polaris system is cleaned and/or decontaminated prior to servicing the equipment, removing it from use, or transporting it for disposal. Refer to the instructions contained in this document and use only those materials specified.

  • Page 47: Biological Agents

    Survey the instrument with an appropriate radioactivity measuring device. Ensure that the survey results are at or below background level. Disposal of IFCs Dispose of used IFCs in accordance with federal, state, regional, and local laws for hazardous waste disposal. Polaris: User Guide...

  • Page 48: Fuse Replacement

    Replace both fuses (8 A, 250 V type, time-lag T fuse), and reinstall the fuse holder. Follow the arrow marks in the fuse holder to match the casing. Plug the instrument into the wall outlet, and toggle the main power switch ON. Polaris: User Guide...

  • Page 49: Appendix C: Safety

    The boxed Polaris system weighs 110 kg (242 lb). If you choose to lift or move Polaris after it has been installed, do not attempt to do so without the assistance of others.

  • Page 50: Instrument Symbols

    Conformity mark Description Indicates conformity with European Union requirements for safety and electromagnetic compatibility. Indicates conformity with North American requirements for safety and electromagnetic compatibility. Polaris: User Guide...

  • Page 51: Electrical Safety

    Electrical Safety WARNING ELECTRICAL HAZARD. DO NOT REMOVE THE COVERS. Electrical shock can result if the Polaris instrument is operated without its protective covers. There are no user serviceable components inside. WARNING ELECTRICAL HAZARD. Plug the system into a properly grounded receptacle with adequate current capacity.
  • Page 52 For technical support visit fluidigm.com/support.

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