Autoradiography data can be loaded, viewed, analyzed, and saved using the same tools VivoQuant uses for in vivo image data. Once loaded into VivoQuant, data can be reoriented, cropped, rescaled, and quantified.
The Autoradiography Calibration tool allows users to calibrate autoradiography data based on known calibration standards in the image.
Getting There
To access the tool, activate to the 3D ROI Tool operator, then go to the Operator tab in the main menu and select Autoradiography Calibration.
Function
Once the tool starts, it will ask questions to establish how it will be used. The first prompt asks for the number of calibration standards being used. This will determine the number of ROIs that populate the 3D ROI Tool.
The second prompt asks if there will be a background ROI used. This influences the calibration curve. In most cases a background ROI is recommended.
Based on the answers to the previous questions, the ROIs will be populated. Now, they need to be manually placed in the correct locations in the image. Use the 3D Paint Mode tools to perform this task. Regions should appear similar to the image below.
Image Calibration
Once the calibration standards have been segmented, click Read Values from Image. The table will display the mean value of each region. Provide the true value of each standard as well as the correct output unit. Also, choose from the available Error Weight techniques.
Once the settings are established, click Run. The plot will populate with the values of the standards and provided values along with the calculated fit line. Changing the settings, particularly the Error Weights, will change the output calibration curve.
To see how the curve changes with different settings, change them and again click Run. Once the curve fits as desired, click Apply to apply the calibration to the data.
