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Calibrating Continuous Flow Chips

Continuous Flow (CF) chips must be calibrated whenever you want to use them with reagents that have not already been calibrated for continuous flow use. CF chip calibration determines the amount of fluid dispensed per unit of time. This tutorial will show you how to calibrate the CF chips for a more consistent flow rate, resulting in a more accurate dispense.

Here, you will learn how to calibrate a new reagent, use the default data for a 1 cP liquid (such as pure water), or even copy any existing calibration data from a similar reagent. The calibration procedure includes inputting some information about your reagent, instrument setup, defining the dispense parameters, preparing the tools needed for calibration, and then dispensing the reagent.

Tip: For quicker calibration, you can import a .CSV file of a 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 Across MANTISes.

Required Tools

• Either one of the following options:

  • The FORMULATRIX Calibration Kit, which includes an SBS footprint microtube rack that works best for the calibration process. If you have updated to MANTIS 4.7.4 or newer, but do not have the kit, please contact support@formulatrix.com.

    OR

  • Four weigh boats.
    
    

  Important: If you do not know the density of your reagent, please prepare one additional microcentrifuge tube or weigh boat for use in defining the liquid density with MANTIS.

• A weighing balance with 0.001 g readability
• A continuous flow chip and tubing harness

Prerequisites

• You are running MANTIS version 4.7.4 or newer.
• You have prepared the desired reagent to be calibrated (or a good surrogate with closely similar density, viscosity and surface tension) and the required tools.
• You have at least one Continuous Flow calibration data in the MANTIS database in order to use the Copy Existing Calibration Data option. Otherwise, this option will be hidden.

To calibrate a Continuous Flow chip:

1. Go to the Tools menu, click Continuous Flow Calibration.

2. In the Continuous Flow Options window, select Continuous Flow Calibration and then click Next.

   

   Continuous Flow Option Window

3. In the Continuous Flow Calibration wizard, select the calibration type from the following options:

   New Calibration

   1. Click New Calibration to create a new calibration for the desired reagent and click Next.

      

      Select New Calibration in the Continuous Flow Calibration Wizard

   2. Select the chip port position to assign your reagent from the Select Chip Port Position list, then click Next.

      Note: When you select the chip port position, the chip's serial number will automatically display in the Chip Serial Number field.

      

      Select the Chip Port Position

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

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

      2. Select the Reagent Viscosity for your reagent. MANTIS CF can support viscosities from 1 to 25 cP. This viscosity value is used for nozzle recommendation and does not impact the calibrated volume.

      3. If you know the density of your reagent, select I know the Reagent Density and type the Density for your reagent in g/mL in the corresponding field. If you do not have the reagent density value and want to measure it during the calibration process, clear the I know the Reagent Density checkbox, if selected.

      4. Select the Nozzle Size of your chip from the available options, such as 0.1 mm - Red, 0.6 mm - Maroon, and so on.

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

         Important: Make sure to select the supported Nozzle Size and Bottle Pressure based on your reagent viscosity to prevent any dripping during dispense. If this warning signappears next to the Nozzle Size or Bottle Pressure field, hover over it to see the recommended bottle pressure and nozzle size values from MANTIS. You can ignore these warnings and continue the calibration, but the calibration may not produce a quality dispense.

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

      

      Data Calibration Has Been Set Up and Ready Proceed

   4. Click Start Calibration to proceed to the following instructions:

   Specify the Calibration Kit.

   Do you have the Calibration Kit from FORMULATRIX?

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

     OR

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

     

   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	Preparing the microcentrifuge tubes: Prepare four empty microcentrifuge tubes to contain the reagent. Place the four tubes in the vial rack in positions 1 through 4, as shown in the image below. Put the Four Empty Microcentrifuge Tubes in the Vial Rack Important: If you have not previously defined the reagent density in the Density field of the wizard, you must place an additional microcentrifuge tube in position 5, as shown in the above image. Click Next to continue. Weigh each empty microcentrifuge tube using the weighing balance. Type the weight value (in milligrams) of each tube under the Empty Weight column, then click Next. Take care not to mix the tubes up. Start the calibration: Place the vial rack with the microcentrifuge tubes on the MANTIS plate holder as shown in the image below, then click Next. Place the Vial Rack and Microcentrifuge Tubes on the MANTIS Plate Holder Dispense the reagent to the four microcentrifuge tubes by clicking the Dispense button. After the dispense is finished, weigh each tube, then type the weight value (in milligrams) in the fields under the Final Weight column. Empty weight subtraction will be done automatically based on the values entered in the step ii. Important: In order for MANTIS to automatically calculate the reagent density, manually add 1 mL of the reagent to the empty tube using a calibrated pipette, then weigh the tube. Type the final weight in milligrams for microcentrifuge tube #5 under the Final Weight column. Click Calculate when you’re done. Then, proceed to step 4.
   Weigh Boats	Preparing the Weigh Boats: Prepare four empty weigh boats to contain the reagent. Label the weigh boats with different numbers (1-4), so that they will not get mixed up. Click Next to continue. Weigh each empty weigh boat using the weighing balance. Type the weight value (in milligrams) of each weigh boat under the Empty Weight column, then click Next. Start the calibration: Place a weigh boat in the A1 position on the MANTIS plate holder, as shown in the picture below. Then, click Next. Put the Weigh Boat in A1 Position on the MANTIS Plate Holder Click the Dispense button to dispense the reagent into the weigh boat. After the dispense is finished, weigh the weigh boat which contains the dispensed reagent, and then type the weight value (in milligrams) in the field under the Final Weight column. Click Next, then repeat steps a-c to dispense the reagent into the other three weigh boats. Click Calculate when you’re done. Then, proceed to step 4. Important: If you did not previously define your reagent density in the Density field of the Calibration wizard, follow the steps below: 1. Prepare and label a fifth weigh boat, then proceed to step ii above 2. Manually add 1 mL of the reagent using a calibrated pipette to the empty weigh boat. Then, weigh the weigh boat and type the weight (mg) in Weigh Boat #5 under the Final Weight column. 3. Click Calculate in order for MANTIS to automatically calculate the reagent density,

   Copy Existing Calibration Data

   1. To calibrate a reagent using the existing calibration data from similar reagent, click Copy Existing Calibration Data, and then click Next.

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

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

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

         

         Copy the Existing Calibration Data

   3. Click Copy Calibration and proceed to step 4.

   Calibration using Default Value

   1. For any reagent with approximately 1 cP viscosity (water-like), you can click Calibration using Default Value, 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 Default Data for 1 cP Viscosity Reagent

   4. Click Apply Calibration to apply the setup value and proceed to step 4.

4. Congratulations! You have successfully calibrated the reagent for a CF chip. You can see the results of the calibration displayed in the wizard. Click Finish when you’re done.

   

   Calibration Is Complete

Important: For the most accurate dispense possible, we recommended recalibrating your reagents each time you change CF chips. To do so, open the Continuous Flow Calibration wizard, select the New Calibration option, then follow the instructions of the New Calibration above. If you recalibrate a reagent, the previous calibration data will be replaced by the new calibration setup.

Related Topics

• Designing Dispenses
• Cleaning the Chips
• Instrument Overview
• Continuous Flow
• Exporting and Importing Continuous Flow Data Across MANTISes

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