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Calibrating Continuous Flow Chips

Continuous Flow (CF) and High Viscosity Continuous Flow (HVCF) chips must be calibrated before you use them with new reagents to determine the amount of fluid dispensed per unit time. Every reagent may have different calibration values with different chips; therefore, it is preferred to do a new calibration for each reagent when used for the first time with any CF chip.

In this section, you will learn how to calibrate a new reagent, use the example calibration data, or even copy any existing calibration data from a similar reagent.

See the table below to help select the appropriate continuous flow chips.

Tip: For quicker calibration, you can import a .CSV file of Continuous Flow calibration data from your computer to the Mantis software. You can also export the calibration data of your calibrated reagents for future reference. For more information, see Exporting and Importing Continuous Flow Data.

Software and Instrument Compatibility

• Mantis software version 5.1 or newer

• Continuous Flow feature license

• Mantis instrument V4

  Note: The standard CF chip is also compatible with Mantis instrument V3.

Required Tools

• Either one of the following options:

  • The FORMULATRIX Calibration Kit with 6 microcentrifuge tubes. If you do not have the calibration kit, please contact support@formulatrix.com.

    OR

  • Six weigh boats
    

• A balance with 0.001 g readability

• A standard CF or HVCF chip and tubing harness

Prerequisites

• You have prepared the reagent to be calibrated (or a good surrogate with closely similar density, viscosity, and surface tension).

  Note: To enable the Copy Existing Calibration Data option, you will need at least one Continuous Flow calibration in the Mantis database.

  

The Copy Existing Calibration Data Option is Active

To calibrate a CF or HVCF chip:

1. Choose from the following options to open the Continuous Flow Calibration wizard:

   • Click the Input button of the uncalibrated reagent in the Input Control Panel. Select Continuous Flow or High Viscosity Continuous Flow from the Chip Type option. Then, click Start New Calibration.

     

     Input Options Window

     Note: If you calibrate the chip from the Input Options window, Mantis automatically defines chip port position, chip serial number, and reagent name.

     OR

   • Go to the Tools menu, and click Continuous Flow Options. Select Continuous Flow Calibration, and then click Next.

     

     Continuous Flow Option Window

2. Select the calibration type from the following options:

   New Calibration

   1. Click New Calibration to calibrate a reagent for the first time, and click Next.

      

      Click New Calibration

   2. Select the chip port position of the assigned reagent. The chip’s serial number will automatically appear in the Chip Serial Number field. Click Next.

      

      Select the Chip Port Position

   3. Specify the calibration setup by following the steps below.

      Important: For HVCF chip calibration, follow the HVCF Chip Calibration Recommendation for accurate results.

      1. Next to Reagent Name, select the reagent you want to calibrate.

      2. Select the Reagent Viscosity for the standard CF chip from 1 to 30 cP. This viscosity value is used for nozzle recommendation and does not impact the calibrated volume.

         Note: If you calibrate the HVCF chip, the Reagent Viscosity will be automatically be set to High Viscosity.

      3. If you know the density of your reagent, select I know the Reagent Density and type the Density in g/mL. If you do not have the reagent density value, uncheck the I know the Reagent Density checkbox.

      4. Select the Nozzle Size from the available options.

      5. Select the appropriate Bottle Pressure value from the following options: 3, 6, 10, 13, and 16 psi.

      6. Select the Bottle Size accordingly: Small (100 mL), Medium (100-250 mL), or Large (250-500 mL). Recommended Bottle Size for HVCF chip is Medium (100-250 mL).

      

      Set the Calibration Data for the Selected Reagent

      Important: If this warning indicator appears next to the Nozzle Size or Bottle Pressure field, hover over it to see the recommended bottle pressure and nozzle size.

      

      Warning Indicators

   4. Click Start Calibration to proceed.

   Specify the Calibration Kit.

   • Click Yes, I have the Calibration Kit if you want to use a tube rack and Eppendorf-style tubes for calibrating. Then, click Next.

     OR

   • Click No, I don’t have the Calibration Kit if you want to use weigh boats. Then, click Next.

     

   Specify the Calibration Kit

   Prepare the Calibration Kit or weigh boats for use in the calibration.

   The table below explains in detail the required preparation for the microcentrifuge tubes and the weigh boats.

   Calibration Tools	Instructions
   Microcentrifuge Tubes	Follow the on-screen instructions to prepare the microcentrifuge tubes on the vial rack. For standard CF chip: If you have not previously defined the reagent density in the Density field, place an additional microcentrifuge tube in position 5, as shown in the image. Put the 5 Empty Microcentrifuge Tubes in the Vial Rack For HVCF chip: If you have not previously defined the reagent density in the Density field, place an additional microcentrifuge tube in position 5, as shown in the image. The software may also prompt you to place an additional microcentrifuge tube in position 6, if necessary. Put the 5 and 1 Additional Empty Microcentrifuge Tubes in the Vial Rack Click Next to continue. Weigh each empty microcentrifuge tube using a balance. Type the weight value (in grams) of each tube in the Empty Weight column, then click Next. Type the Weight Value in Each Empty Weight Column Start the calibration: Place the tube rack on the Mantis deck as seen below. Then, click Next. Vial Rack for CF Chip Calibration Vial Rack for HVCF Chip Calibration Define the Priming Pulse Duration value for priming. Then, click Start Prime to begin priming the reagent. Click Stop Prime when an uninterrupted stream of reagent leaves the chip nozzle. Then, click Next. Priming the Chip Click the Dispense button to dispense the reagent. After the dispense is finished, weigh each tube, then type the weight value (in grams) in each Final Weight field. Click Next. Type the Weight Value in Each Final Weight Column OPTIONAL: To calculate the reagent density: Manually add 1 mL of the reagent to the empty tube in position 5 using a syringe or calibrated pipette. Weigh the tube, and type the final weight (in grams) for microcentrifuge tube #5 under the Final Weight column. See the following options: For the standard CF chip, click Calculate. For the HVCF chip, click Next. If prompted for an additional measurement by the software, continue to step e. Define the Reagent Density OPTIONAL for HVCF chip: Follow the on-screen instructions to fill and weigh microcentrifuge tube #6. Once it’s finished, click Calculate. Define the Additional Measurement Continue to step 3.
   Weigh Boats	Follow the on-screen instructions to prepare the Weigh Boats. For standard CF chip: If you have not previously defined the reagent density prepare 5 weigh boats, as shown in the image. Prepare 5 Weight Boats For HVCF chip: If you have not previously defined the reagent density, prepare 6 weigh boats. The software may also prompt you to run an additional measurement in weigh boat #6, if necessary. Prepare 6 Weight Boats Weigh each empty weigh boat using the balance. Type the value (in grams) in each Empty Weight field. Click Next. Type the Weight Value in each Empty Weight Column Start the calibration: Place a weigh boat in the A1 position on the Mantis deck, as seen below. Then, click Next. Place the Weigh Boat in A1 Position on the Mantis Deck Define the Priming Pulse Duration value for priming. Then, click Start Prime to begin priming the reagent. Click Stop Prime when an uninterrupted stream of reagent leaves the chip nozzle. Then, click Next. Priming the Chip Click the Dispense button to dispense the reagent into each weigh boat. Click the Dispense Button After the dispense is finished, weigh the full weigh boat and enter the value (in grams) in the field under the Final Weight column. Type the Final Weight Click Next to define the other three weigh boats. OPTIONAL: If you did not previously define your reagent density, follow these steps: Prepare and label a fifth weigh boat, then proceed to step 2 above. Add 1 mL of the reagent using a calibrated pipette to the empty weight boat. Then, weigh the weigh boat and type the weight (in grams) in Weigh Boat #5 under the Final Weight column. See the following options: For standard CF chip, click Calculate. For HVCF chip, click Next. If prompted for an additional measurement by the software, continue to step g. OPTIONAL for HVCF chip: Follow the on-screen instructions to fill and weigh weigh boat #6. Once it’s finished, click Calculate. Define the Additional Measurement Continue to step 3.

   Copy Existing Calibration Data

   1. Select Copy Existing Calibration Data, and then click Next.

      

      Select Copy Existing Calibration Data

   2. Specify the calibration setup by following the steps below.

      1. Define the Reagent Name. If you copy calibration data from the Input Options window, the reagent name will be automatically generated.

      2. Select the desired calibration data from the Select Existing Calibration Data list. The copied calibration data will be automatically displayed in each respective field.

      3. Click Copy Calibration.

         

         The Existing Calibration Data

      4. Click Finish.

   Use Example Calibration

   Select this option to calibrate the CF chip for use with any reagent with approximately 1 cP viscosity (water-like).

   Important: This option is unavailable for HVCF chip calibration.

   1. Select Use Example Calibration, then click Next.

   2. At this stage, the Reagent Viscosity field has been automatically assigned to 1 cP.

   3. Define the nozzle size, bottle pressure, and bottle size in the provided fields.

      

      Use Example Data for 1 cP Viscosity Reagent

   4. Click Apply Calibration to apply the setup value.

   5. Click Finish.

3. Calibration completed! The calibration results will be generated in the last page of the wizard. Click Finish.

   

   The Calibration Result

Note: If you calibrate a CF chip from the Input Options window, the calibration data will automatically be generated under the Calibration Data section. Click OK to save the changes.

The CF Calibration Data

HVCF Chip Calibration Recommendation

The following tables are for general reference. The calibration results may vary depending on the chip lifetime, nozzle size, and pressure used during calibration.

Important: If you dispense high-viscosity reagents using an HVCF chip, maintain consistent room temperature and humidity. When changing the chips, recalibrate your reagents using the New Calibration option. The new setup will automatically replace the previous calibration data.

Related Topics

• Designing a Dispense
• Washing the Mantis Chips
• Hardware Overview
• Continuous Flow
• Exporting and Importing Continuous Flow Data

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