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 can 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 as well as instrument setup and defining the dispense parameters, preparing the tools needed for calibration, and then dispensing the reagent.

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 4.3 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 4.3 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.

Continuous Flow Calibration Wizard

To calibrate a Continuous Flow chip:

Go to the Tools menu, click Continuous Flow Calibration.
Next to Reagent Name, select the reagent you want to calibrate.
Select the Calibration Type from the following options:
- To create a new calibration for the desired reagent, click New Calibration, then proceed to step 4.
  
  OR
- For any reagent with approximately 1 cP viscosity (water-like), you can click Use Default Data for 1 cP Liquid, then define the nozzle size, bottle pressure, and bottle size (see steps 6-8 below for more information). Click Apply Calibration to apply the setup value. When the calibration has been successfully completed, click Finish. Note that the accuracy may suffer if the viscosity is significantly different from 1 cP.
  
  OR
- To calibrate the desired reagent using existing calibration data from similar reagent, click Copy Existing Calibration Data, choose the desired calibration data from the Select Existing Calibration Data list, then click Copy Calibration. When the calibration has been successfully completed, click Finish.
Select the Reagent Viscosity for your reagent. MANTIS CF can support viscosities from 1 to 25 cP.
If you know the density of your reagent, select I know the Reagent Density and type the Density for your reagent in g/L. 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.
Select the Nozzle Size of your chip from the available options, such as 0.1 mm - Red, 0.6 mm - Brown, and so on.
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 sign appears 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 result might not be accurate.
Select the Bottle Size accordingly: Small (100 mL), Medium (100-250 mL), or Large (250-500 mL).

Click Start Calibration to proceed to the following instructions:

C. Prepare the Calibration Kit or weigh boats for use in the calibration. The table below explains in detail the required preparation for either the microcentrifuge tubes or 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: To let MANTIS automatically calculate the reagent density, add 1 mL of the reagent to the empty tube manually using a trusted 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 10.
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 10. Important: If you did not previously define your reagent density in the Density field of the wizard, follow the steps below: 1. Prepare and label a fifth weigh boat, then proceed to step ii above 2. Add 1 mL of the reagent using a trusted pipette to the empty weigh boat, then weigh the weigh boat and type the weight in milligrams in Weigh Boat #5 under the Final Weight column. 3. Click Calculate to let MANTIS automatically calculate the reagent density,

Calibration Tools

Instructions

Microcentrifuge Tubes

Preparing the microcentrifuge tubes:
1. Prepare four empty microcentrifuge tubes to contain the reagent.
2. 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.
3. 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:
1. 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
2. Dispense the reagent to the four microcentrifuge tubes by clicking the Dispense button.
3. 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: To let MANTIS automatically calculate the reagent density, add 1 mL of the reagent to the empty tube manually using a trusted pipette, then weigh the tube. Type the final weight in milligrams for microcentrifuge tube #5 under the Final Weight column.
4. Click Calculate when you’re done. Then, proceed to step 10.

Weigh Boats

Preparing the Weigh Boats:
1. Prepare four empty weigh boats to contain the reagent.
2. Label the weigh boats with different numbers (1-4), so that they will not get mixed up.
3. 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:
1. 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
2. Click the Dispense button to dispense the reagent into the weigh boat.
3. 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.
4. Click Next, then repeat steps a-c to dispense the reagent into the other three weigh boats.
5. Click Calculate when you’re done. Then, proceed to step 10.
Important: If you did not previously define your reagent density in the Density field of the wizard, follow the steps below:
1. Prepare and label a fifth weigh boat, then proceed to step ii above
2. Add 1 mL of the reagent using a trusted pipette to the empty weigh boat, then weigh the weigh boat and type the weight in milligrams in Weigh Boat #5 under the Final Weight column.
3. Click Calculate to let MANTIS automatically calculate the reagent density,

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, it is recommended to recalibrate your reagents each time you change between CF chips. To do so, open the Continuous Flow Calibration wizard and select the desired reagent that has previously been calibrated. Select New Calibration as the Calibration Type option, click the Re-calibrate button, then follow steps 9-10 in the procedure above. If you recalibrate a reagent, the previous calibration data will be replaced by the new calibration setup.

Calibrating Continuous Flow Chips

Required Tools

Prerequisites

To calibrate a Continuous Flow chip:

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