How do I Collect Fluorescence Data from an FLR/FLD using a Generic Instrument in ASTRA - WKB319060
OBJECTIVE or GOAL
To configure ASTRA to collect and report analog fluorescence data from an FLR/FLD detector using a generic instrument setup, enabling integration of fluorescence signals into SEC-MALS or FFF-MALS experiments.
ENVIRONMENT
- ASTRA
- DYNAMICS
- Instrument: FLR/FLD (Fluorescence Detector)
- Instrument: MALS (Multi-Angle Light Scattering)
- Software: ASTRA
- Instrument Configuration: Generic Viscometer
PROCEDURE
Open your ASTRA method and activate Experiment Builder Mode. Add a Generic Viscometer instrument and an Aux Channel Connection in the Configuration section. Set the Source Device to the Generic Viscometer and the Destination Device to your MALS instrument, then designate the appropriate AUX Channel. Save the method and exit Experiment Builder Mode. Use this method to collect FLD signals as analog data labeled “VIS” within ASTRA.
ADDITIONAL INFORMATION
Collecting Fluorescence Data from an FLR/FLD using a Generic Instrument in ASTRA
If a fluorescence detector (FLR/FLD) is used to collect data as a part of your SEC-MALS or FFF-MALS experiment, you can set up your ASTRA method to collect this data (alongside other data such as UV, MALS, RI, etc.) for your samples. While fluorescence data cannot be used quantitatively in the context of light scattering measurements, it can be incredibly helpful to have this data accessible within an ASTRA experiment file as an additional means of characterization of the sample. Incorporating this data within ASTRA additionally streamlines data reporting, enabling reporting of fluorescence data as a part of your SEC-MALS or FFF-MALS experiments.
This guide article will describe the procedure to modify your ASTRA method to use a generic instrument as a means to collect and report analog data from your fluorescence detector.
Setting up a Generic Instrument in an ASTRA Method
In ASTRA, open the ASTRA method you would like to modify by navigating to File → New → Experiment from Method.Activate Experiment Builder Mode by navigating to System → Preferences → Experiment Builder Mode.Double click the Configuration of the method and expand Details. Here we will add both an instrument and an aux connection.To add the Generic Instrument, click Browse... next to Instruments → Add. From the topmost folder, navigate to System → Configurations → Viscometry → Generic Viscometer and add the generic VI instrument.To add the Aux Connection, click Browse... next to Connection → Add. From the topmost folder, navigate to System and add the aux channel connection.You should now see the Generic Viscometer and a new Aux Channel Connection, click on the Aux Channel Connection to open it up.
- Designate the Source Device as the Generic Viscometer, and the Destination Device as your MALS instrument (or alternatively whichever instrument is receiving the analog signal). Finally, designate the Destination AUX Channel. If you are unsure which channel to choose, this can be configured later (see note on this below: Checking the Destination AUX Channel).
- Click OK to save these changes.
- Close Experiment Builder Mode by navigating again to System → Preferences → Experiment Builder Mode.
- Save the method by right-clicking the experiment name and selecting Save as Method.
You can now use this method to collect your FLD signal as an analog signal alongside your data in ASTRA. Note that this data will be labeled as "VIS" for viscometer but the data will be that exported by your FLR/FLD instrument.
Checking the Destination AUX Channel
If you are unsure which Aux channel to designate for the generic instrument, this will correspond to the analog connection made on the rear panel of your Wyatt instrument (typically done with your MALS instrument). For more information on making these analog connections and example for different HPLCs, see the corresponding HPLC Connection Guide (TN3600 series).
To confirm which Aux connection is being used or to troubleshoot the connection, open an ASTRA experiment file that was configured as described above and used to that will show a fluorescence signal). Navigate to Experiment → Graph → Add Custom Plot:
In the window that opens, designate original data and click the >>> button to carry this over to the Included section.
Next, check the box for Raw Aux Input Data, as shown below, and click OK.
In the Results section of the experiment file, there is now a new graph, double-click this graph to open it:
Use the checkboxes to view/hide the data obtained from the aux connections of the instrument. Each trace is labeled with the corresponding channel number. All aux signals are always connected by your Wyatt instruments, regardless of if this graph or if the method described above is set up. Note the channel number for the trace that corresponds to your FLD signal, this will be the Destination Channel as described above.