Nicoya OpenSPR Operation Manual
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Nicoya OpenSPR Operation Manual

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NICOYA LIFESCIENCES
1-CHANNEL OPERATION MANUAL V4.1.01
Released: January 18, 2018 (V4.1.01)
support@nicoyalife.com

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  • Page 1 NICOYA LIFESCIENCES 1-CHANNEL OPERATION MANUAL V4.1.01 Released: January 18, 2018 (V4.1.01) support@nicoyalife.com...

  • Page 2: Table Of Contents

    Before any steps are taken ..................7 1.1.2 Standard Equipment Supplies ................... 7 1.1.3 Setting up the OpenSPR .................... 8 1.1.4 Connecting and Turning on the OpenSPR............... 10 OpenSPR Software Setup ....................11 1.2.1 Minimum Computer Requirements ................ 11 1.2.2 Installing OpenSPR Software .................. 11 TraceDrawer Software Setup ..................
  • Page 3 Preparation of Reagents ....................55 Standby Mode ........................ 55 Instrument Shutdown ....................55 Recommended Cleaning ....................56 3.4.1 Regular Cleaning ..................... 56 3.4.2 Advanced Cleaning ....................57 SYSTEM DETAILS ........................59 OpenSPR Hardware ......................59 OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 4: Safety And Preparation For Use

    SAFETY AND PREPARATION FOR USE Line Voltage The OpenSPR™ operates using a UL certified Class 2 power supply with an input of 100-240 V, ~50-60 Hz and 2.25 A maximum. Only use the power supply provided with your instrument. Operate Only with Covers in Place To avoid personal injury, do not remove the product covers or panels.
  • Page 5 4. Ensure proper ventilation while in use. 5. Do not operate unattended. 6. OpenSPR is not waterproof – do not spill liquids onto it and be very careful of leaks. 7. Do not operate the system in the presence of strong magnetic fields.
  • Page 6 17. Turn off the OpenSPR immediately if a leak is detected. 18. Only use blunt-end injection tips in the OpenSPR. Using sharp needle tips will damage the injection port, and replacement will be at the user’s expense. This information is based on our present knowledge. However, this does not constitute a guarantee for any specific product features and shall not establish a legally valid contractual relationship.

  • Page 7: Getting Started

    Prior to setting up your OpenSPR, ensure your laboratory space is setup with the below requirements: • A clean and level lab bench with enough space to fit both the OpenSPR and laptop computer (3.5 ft or 1 m of bench space is recommended) •...

  • Page 8: Setting Up The Openspr

    Screwdriver (for servicing) 1.1.3 Setting up the OpenSPR 1. Remove the OpenSPR Instrument from the box, and place on a clean, level surface. 2. Remove the Power Cable and USB Cable from the bottle tray of the instrument (right- hand side) and remove all additional packaging tape present on the instrument.
  • Page 9 9. Insert the bottles into the tray by tilting the bottle such that the base of the bottle fits into the tray first, followed by the top [Figure 1.3]. Figure 1.3 Left: Insert the bottles into the tray base-first and tilted. Right: Proper positioning of all bottles installed in the OpenSPR. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 10: Connecting And Turning On The Openspr

    Figure 1.4 USB and power cable ports on the back of the OpenSPR. Plug one end of the USB Cable into the back of the OpenSPR [Figure 1.4: USB port for computer connection] and connect the other to the computer.

  • Page 11: Openspr Software Setup

    TraceDrawer Analysis Software, you are all set to go! Skip down to Activation of TraceDrawer Analysis software section. If you did not purchase a laptop with your OpenSPR, the USB drive in the Accessories Box will contain the installer for both the OpenSPR and TraceDrawer software.

  • Page 12: Tracedrawer Software Setup

    1.3.2 TraceDrawer License Activation Locate your TraceDrawer License Key. A print-out containing your license key is provided within the folder of the OpenSPR Accessories box. Run the TraceDrawer program from the desktop shortcut. Click “Add Legacy License Key” in the window prompt.
  • Page 13 Tip: It is recommended to create a local file on your computer containing this license key. When Windows Updates occur, you may be prompted to re-enter the TraceDrawer license key. Figure 1.7 TraceDrawer home screen once activated with license key. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 14: Standard Operating Procedures

    Prior to starting your experiment, the fluidic setup must be in place for proper function of the OpenSPR. This includes having the Waste Bottle, DI Water Bottle, and Buffer Bottles filled to the proper levels with tubes installed correctly. Details for each bottle and tubing setup are provided below.
  • Page 15 • Gently tighten the fitting on the tube such that it is secured into place. • Place the bottle onto the tray of the OpenSPR next to the Waste Bottle [Figure 2.2]. Buffer Bottle 1 and 2: 250 mL square glass bottles containing a tube fitting •...

  • Page 16: Optical Setup

    OpenSPR Sensor Chips (red sensor spot) [Figure 2.3]. The 1-Channel OpenSPR is not compatible with Nicoya’s High Sensitivity Sensors as a different optical setup is required. For use of the High Sensitivity Sensors upgrade packages are available – please contact a Customer Success scientist for more details.

  • Page 17: Beginning A Test

    2.2 Beginning a Test Prior to beginning a test for the first time after receiving and unpacking your OpenSPR Instrument, ensure you complete a hands-on training with a Customer Success Scientist at Nicoya Lifesciences. Verify the fluidic setup is complete (Section 2.1.2).

  • Page 18: Instrument Setup

    OpenSPR, then re-establish the USB connection. If the software is still unable to connect, restart the software. Tip: to power off the OpenSPR, hold the power button down for 10 seconds until the power LED turns off Click “Start” to enter the software. The software will walk you through the Instrument Setup, and Sensor Setup procedures prior to collecting your data –...

  • Page 19: Naming Your Running Buffers

    Bottle 1 and Bottle 2 [Figure 2.9]. You will also be given the option to input the names of the running buffer during the Priming process as well (Section 2.3.2). Note that the DI Water and Waste bottle designations cannot be changed. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 20: Step 1: Fluidic Priming

    The completion of this step is not required for running a test but is recommended for your experimental records. 2.3.2 Step 1: Fluidic Priming Priming of the OpenSPR will fill the pumps and tubing with the running buffer to be used for the test. It is necessary to perform priming in the below scenarios:...
  • Page 21 1. To setup the instrument for priming, ensure that a chip (either a blank chip or a sensor chip) is installed in the sensor holder and docked in the system. The OpenSPR will come pre-installed with a blank chip for your convenience upon receiving. In the prompt, you will be able to undock the sensor stage in order to insert or change a chip if necessary [Figure 2.11].
  • Page 22 3 mL of the running buffer used for priming (taken from the separate buffer bottle set aside for this purpose). Figure 2.14 Instrument priming progress bar. 5. When the priming process is complete, the priming status in the software will be updated accordingly. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 23: Step 2: Optical References

    The date and time when the previous references were acquired will be displayed on the main screen [Figure 2.15]. Figure 2.15 Date and time previous references were acquired as well as expected reference for 1-Channel OpenSPR. It is only recommended to take new references in the below scenarios:...
  • Page 24 The process for taking new optical references is detailed below: 1. To be able to take the references, a clean blank chip must be installed in the OpenSPR [Figure 2.17]. To install a blank chip into the OpenSPR, follow the directions below.
  • Page 25 6. Install the holder containing the blank chip into the sensor stage, close the lid of the OpenSPR, then dock the stage using the button available in the window. 7. Once the blank chip is fully installed into the OpenSPR, select “Next” in the window [Figure 2.17] to proceed.
  • Page 26 Figure 2.20 Optical references bubble inspection and removal. Figure 2.21 Optical references bubble removal in progress. 10. When the optical path is cleared of all bubbles, close the lid of the OpenSPR and select “Next” to begin taking the optical references. The process will take approximately 2 minutes to complete and the progress will be displayed [Figure 2.22].

  • Page 27: Sensor Setup

    * Note that the status of each of the processes shown and the plots of the absorbance graphs will be different depending on the instrument state. 2.4.1 Step 3: Install Sensor Chip To install a new sensor for your test, click the “Load Sensor” button [Figure 2.24]. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 28 [Figure 2.25]. To place a new sensor into the instrument, ensure the lid of the OpenSPR is closed and select “Load New Sensor”. At this time the sensor stage will automatically undock for you.
  • Page 29 Figure 2.26 Prompt instructing user to remove existing sensor or chip from the system. 3. Once the stage is fully undocked and stationary, open the lid of the OpenSPR and remove the sensor holder containing the existing chip from the system. To do this, lift the tabs of the sensor holder straight out of the stage [Figure 2.27].
  • Page 30 Figure 2.30, you will be viewing the front sensor surface. Figure 2.30 With the sensor chip in the vertical position, when the cut corner is in the bottom right, you are viewing the front sensor surface. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 31 9. Place the holder containing the sensor into the docking stage. Ensure it is fully seated within the docking stage. 10. Close the lid of the OpenSPR, then click “Dock” to allow the sensor to be automatically docked into place, creating a seal with the flow cell.
  • Page 32 If no leaking is present, select “Next” to proceed onto the bubble removal step (Section 2.4.2). Once a sensor is loaded into the OpenSPR, an absorbance peak should be seen in the graph on the main screen [Figure 2.34]. The center position of the absorbance peak should be near approximately 550 nm.

  • Page 33: Step 4: Bubble Removal And Prevention

    1. To remove bubbles, a solution of 80% isopropanol will be used. The software will prompt you to load this solution into the injection port – at least 150 µL volume is recommended [Figure 2.36]. The solution should be introduced into the OpenSPR using a designated syringe with a blunt-end injection tip.
  • Page 34 4. Once the flow cell channels are clear, click “Finish” to end the process. The bubble removal status will be updated accordingly on the main screen. 5. Prior to moving forward with your experiment, rinse the injection port with at least 0.5 mL of running buffer. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 35: Running An Experiment

    Upon starting a test, you will be introduced to the main test interface [Figure 2.41]. This interface will allow you to control the various aspects of your OpenSPR instrument. Each aspect will be described in detail in the sections below and how they relate to your experiment.
  • Page 36 Approximately 4 points per second for each channel is being acquired. Figure 2.42 Initial real-time data streaming into the response graph. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 37: Changing The Running Buffer

    2.5.3 Changing the Running Buffer During an SPR experiment, in some scenarios a change of buffer may be needed. The OpenSPR contains an automated process for changing the running buffer, which can be accessed by selecting “Change Buffer” in the menu bar [Figure 2.43].

  • Page 38: Flow Rate Controls

    OpenSPR is flushed. Figure 2.46 Example response graph of a buffer change to a higher refractive index running buffer.

  • Page 39: Injections

    ⁄ �������� �������� (μL min ) The standard sample loop that comes installed in the OpenSPR is 100 µL in volume. For different injection types and instances in your experiment, it can be advantageous to use different flow rates. A table of example scenarios and recommended flow rates is provided below: Table 3 Recommended typical flow rates for different injection types and experimental scenarios.
  • Page 40 OpenSPR with your sample [Figure 2.48]. Figure 2.48 Example of loading sample into disposable plastic syringe with air gap. Glass syringes (SYR-G) are 250 µL in volume and come installed with a gastight blunt-end injection tip which does not contain a dead volume (replacement needles are also available, RN-G-6).
  • Page 41 Before removing the syringe from the injection port, wait at least 5 seconds with the syringe stationary for the fluidic pressure to stabilize in the sample loop. Figure 2.50 Glass syringe for 150 µL sample volume properly seated into OpenSPR. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 42 4. Click “Inject” in the software to introduce your sample over the sensor [Figure 2.51]. At this time, the OpenSPR will automatically introduce your sample loaded into the sample loop into the running buffer flow to be exposed over the sensor. Other instrument functions will be disabled in the software during this time until the injection is complete.

  • Page 43: Ligand Immobilization Wizard

    6. Only after the injection is complete and the baseline has settled should the user move onto the next injection, repeating this procedure. 2.5.6 Ligand Immobilization Wizard The OpenSPR software contains a built-in ligand immobilization wizard which walks the user though the recommended procedures for immobilization of a ligand onto the various sensor chemistries available.
  • Page 44 Once an injection is complete, the user can click “Next” to move onto the following step. Prior to performing the subsequent injection, it is recommended that the user waits until the baseline stabilizes from the previous injection. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 45 If not, it is recommended that the user repeats another injection of the ligand. If the ligand immobilization level achieved is significantly lower than the target, the user should attempt to increase the concentration of the ligand used. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 46: Graph Settings

    Zeroing the graph is recommended at any stage in the experiment where a new baseline is established, and the user wants to make a relative comparison for subsequent injections. An example of this scenario is after the ligand immobilization step, and before injection of analyte for analysis. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 47 The response graph axes will autoscale by default as the data points are collected and the signal changes. However, it can be desirable to zoom in or out on certain areas of the graph as the test progresses. The various functions of manually adjusting the graph are described below: OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 48: Advanced Controls

    Namely, the user will have to ability to start and stop the flow in the OpenSPR, and when the flow is stopped, the user can then undock and dock the sensor stage. Typically, these controls should only be used if leaking arises from the flow cell during a test.

  • Page 49: Shutdown Procedures

    2.6.2. Shutdown the instrument: This option should be selected if the user does not plan to use the instrument in the near future and would like to shut it down. This will guide the user OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 50: Standby Mode

    Section 2.6.3. 2.6.2 Standby Mode When the user has ended testing for the day, it is important to place the OpenSPR into standby mode for proper maintenance of the OpenSPR. Standby mode will set up the instrument under a constant low flow rate (5 µL/min) which will prevent any buildup or blockages in the fluidic lines.

  • Page 51: Instrument Shutdown

    The status of the instrument will be displayed, and the instrument can be left unattended. Important: Do not shut down the software or the OpenSPR instrument while in standby mode. 2.6.3 Instrument Shutdown Prior to shutting down the instrument, it is important to thoroughly rinse and sterilize the tubing to maintain proper performance of the OpenSPR.
  • Page 52 [Figure 2.72]. To do this, place all 3 inlet tubes into a bottle containing 80% isopropanol (multiple bottles can be used as necessary, so long as all inlet tubes are submerged). Once all inlet tubes are submerged, click “Next” to begin the sterilization step. OpenSPR 1-Channel Operation Manual V4.1.01...
  • Page 53 8. Once the shutdown preparation processes are complete, a final dialogue box will appear [Figure 2.74]. Clicking “OK” in this dialogue will safely close the software and power off the OpenSPR instrument automatically. Figure 2.74 Shutdown process complete. OpenSPR 1-Channel Operation Manual V4.1.01...

  • Page 54: Data Output Files

    Documents\OpenSPR\TestResults\ and will have the name given at the beginning of the test (Section 2.5.1). An example of contents of the test folder is shown below [Figure 2.75]. Figure 2.75 Data output folders and files created from a test.

  • Page 55: Maintenance And Cleaning

    3.3 Instrument Shutdown If the OpenSPR instrument is not to be used for over 7 days before the next experiment, it is recommended to perform a full shutdown.

  • Page 56: Recommended Cleaning

    DI water is present in a separate accessible container. Empty the Waste Bottle before commencing. 2. Begin the prime sequence and load a blank into the OpenSPR sample holder. Continue to prime the OpenSPR with DI Water from Buffer Bottle 3. Allow the flow rate to continue at the default speed of 200 µL/min.

  • Page 57: Advanced Cleaning

    DI water is present in a separate accessible container. Empty the Waste Bottle before commencing. 2. Begin the prime sequence and load a blank into the OpenSPR sample holder. Continue to prime the OpenSPR with DI Water from Buffer Bottle 3. Allow the flow rate to continue at the default speed of 200 µL/min.
  • Page 58 8. Rinse the injection port with 2 mL of 0.2 M sodium bicarbonate. While the solution is still within the OpenSPR, perform a bubble removal injection of this solution. 9. Once the bubble removal sequence with the 0.2 M sodium bicarbonate is complete, rinse the injection port with 3 mL of DI water.

  • Page 59: System Details

    To power off the OpenSPR, hold the power button down for 10 seconds until the lights of the OpenSPR turn off.
  • Page 60 Figure 4.2 OpenSPR injection port and power button. Opening the lid of the OpenSPR box will expose the optics system, as well as the fluidics system which interfaces with the sensor chip [Figure 4.3]. Figure 4.3 Optical and fluidic system which interfaces with the sensor chip.
  • Page 61 OpenSPR 1-Channel Operation Manual V4.1.01...

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