How to run the protocol adaptation wizard and rename protocols

Background

Adapted protocols are created by the user when the standard gates do not fit their cell population correctly. Some of these cases can include smaller cell size, multiple cell populations, or excess cell debris, to name a few. Below are the steps to create a new adapted protocol or to rename an existing protocol. Please note that there are differences when performing either of these procedures on an instrument with and without 21 CFR Part 11 enabled.

How to run the protocol adaptation wizard

Non-21 CFR Part 11 enabled instrument:

  • With the NucleoView software open, right click the data set you wish to use as a template and select “Start Protocol Adaptation Wizard.”
  • The Protocol Adaptation Wizard should pop up. Step 1 ensures you have the correct protocol selected, (viability and cell count assay, viability and cell count assay Via 2, Aggregated cells assay, etc.) and ensures you have selected the correct image file when you right clicked in the previous step.

Click on the “Step 2” tab. Step 2 is where we select the proper gating. In some cases, the adapted gates do not automatically load. If this is the case, right click the grey space to the left of the gating windows and select “Reload Post Processing.”

Upon clicking the reload post processing option, two confirmation windows will pop up one after the other. Select “No” on both options to load in your custom gating.

Left click in the gray space to the left of your data to turn it blue.

Click on the “Step 3” tab. Step 3 allows you to customize the volume of your samples, set default dilution values, and the information given on the main screen after running a sample. The default is shown below, but you can change it to live cells/ml, dead cells/ ml, etc.

  • Click on the “Step 4” tab. Step 4 allows you to name the protocol. Once a name has been decided upon please click “Save” and then “Finish.”
  • The protocol name should be found in the “Protocol” box of the NucleoView software.

21 CFR Part 11 enabled instrument:

  • Note: adapted protocols cannot be created on a 21 CFR Part 11 instrument. The adapted protocol must be created on a separate computer then imported onto the 21 CFR Part 11-enabled instrument by a NucleoViewAdmin account.
  • Follow instructions as above until you get to Step 4.
  • To save the protocol for use on a 21 CFR Part 11 instrument, you must select “Save and Lock” (lower left corner) to have the protocol loaded onto that instrument.

To rename an adapted protocol (non-21 CFR Part 11 instrument):

  • Follow instructions as above until you get to Step 4.
  • Change the name of the file and click “Save” then “Finish.”

To rename an adapted protocol (21 CFR Part 11 instrument):

  • Note: adapted protocols cannot be changed on a 21 CFR Part 11 instrument. The adapted protocol must be renamed on a separate computer then imported (by a NucleoViewAdmin account) onto the 21 CFR Part 11-enabled instrument.
  • Follow instructions as above until you get to Step 4.
  • Click “Unlock” protocol. You cannot rename a locked protocol.
  • Change the name of the file and click “Save and Lock” then “Finish.”
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