Software Workflow - Slow Kinetics
This tutorial serves as a guide for a Slow Kinetics experimental workflow on Dianthus α, starting from creating a new experiment in DI.Control, running the acquisition, and exporting data, through to analyzing results in the NanoTemper Hub.
Note: Slow Kinetics is a measurement mode that can be unlocked for Dianthus instruments equipped with Spectral Shift optics. NT.23 instruments are not capable of performing Slow Kinetics measurements, but can be physically upgraded to enable this functionality.
1. Create a New Experiment in DI.Control
View: New Experiment Interface (Figure 1)
Purpose: Start a new Slow Kinetics experiment and define the well layout for the plate.
When you open DI.Control, the New Experiment screen is displayed. You have two options to populate the well information:
Quick Start: Click the Quick Start button to configure the wells manually, one by one.
Excel Import: Click the Excel Import button to load a pre-filled spreadsheet that automatically populates all well information. This option is recommended when you have many conditions to set up.
Once you have selected your preferred method and filled in the well information, proceed to the main acquisition setup screen.
2. Configure the Slow Kinetics Acquisition
View: Main Display Window - Slow Kinetics Setup (Figure 2)
Purpose: Define the acquisition parameters, including the number of cycles, the duration of each cycle, and an optional delay before starting.
On this screen, configure the following parameters:
Number of Cycles: Enter the total number of measurement cycles you want to perform (for example, enter 10 for ten repeated measurements).
Cycle Duration: Set the time duration for each individual cycle. This determines how long each measurement lasts.
Delay Before Start: If you need to wait before the acquisition begins (for example, to allow sample equilibration), enter a delay time. Leave this at 0 if no delay is required.
Verify that all values are correctly entered and that the number of cycles and cycle duration match your experimental protocol.
Once you are satisfied with the settings, add the experiment to the queue and click the Start button to begin the acquisition.
3. Export the Data
View: Plate Mapping View, after acquisition is complete (Figure 2, circled in red)
Purpose: Export the acquired data in JSON format for analysis in the App Hub.
When the acquisition is finished, navigate to the Plate Mapping view. Locate the Export JSON button in the top right corner of the screen (circled in Figure 2). Click it to save your data as a JSON file.
Make a note of where the file is saved, as you will need it in the following steps.
4. Log into the App Hub
View: NanoTemper App Hub - Login and Application Selection (Figure 3)
Purpose: Access the online analysis platform and select the Slow Kinetics Analysis application.
Open your web browser and go to https://hub.nanotempertech.com/apps.
Log in with your credentials.
From the list of available applications, select Slow Kinetics Analysis.
5. Load the JSON Data File
View: File Upload Screen (Figure 4)
Purpose: Import the JSON file exported from DI.Control into the App Hub for analysis.
You can load your file in one of two ways:
Drag and drop the JSON file directly into the upload area.
Click the upload area to browse and locate the file on your computer.
Wait for the upload to complete before proceeding to the next step.
6. Select the Analysis Type
View: Analysis Selection Screen (Figure 5)
Purpose: Choose the appropriate analysis method to process your Slow Kinetics data.
The following analysis types are available:
| Analysis Type | Description |
|---|---|
| Covalent | Select this for covalent binding analysis. |
| kon / koff | Select this to determine association and dissociation rate constants. |
| Affinity over time | Select this to display the dissociation constant (Kd) as a function of time. |
Each option includes associated normalization settings and fitting models. Choose the one that best matches your experimental design.
After selecting your analysis type, click the corresponding button to proceed with data processing.
7. Review the Results
View: Results Display Screen (Figure 6 and Figure 7)
Purpose: Examine the processed data, view fitted curves, and manage your analyses.
The Results screen is organized into the following panels:
Center panel: Displays the main output graphs and fitting results.
Right panel: Shows the ligands and conditions used in the experiment.
Top tabs: Allow you to switch between the analysis results and the raw data view.
Left panel: Provides navigation. Click Overview to see all previous analyses, or create a new analysis from here.
You can rename an analysis at any time by clicking on its title. This helps you stay organized when working with multiple experiments.