Pseudo-Absorbance Program

The Pseudo-Absorbance Program is the first processing step in the characterization and quantification of Analytical Buoyancy Density Equilibrium (ABDE) experiments used to determine AAV capsid loading. In ABDE, solutes are separated according to their buoyant density. This module initializes the data by converting raw intensity scans into pseudo-absorbance data through reference subtraction and baseline correction. Reference scans are defined and subtracted from the sample scans, and the resulting corrected data are saved with the prefix PSAB-. Reference scans used by this module must be acquired from the Reference channel (B) of the same centerpiece.

ABDE

PsAP Process:

1. Click Import Run and load ABDE dataset using a Load Run Data Dialog.

2. Assign the reference channel in the Reference pulldown menu. The reference will automatically be subtracted. Select which Triple to save by clicking the Save checkbox.

3. Ensure the subtraction is saved by apply. Edit name and save PABS- auc file in $HOME/ultrascan3/imports.

Pseudo-Absorbance Conversion

Pseudo-Absorbance Functions:

Database	Select to specify data input from the database.
Local Disk	Select to specify data input from local disk or the database.
Import Run	Click here import the dataset using Load Data Dialog, select a run ID for the data set to load.
Load Reference Scan	Load a reference scan from a different dataset if no Reference Scan was included.
Reset	Reset all parameters to defaults.
Run ID	the Run identifier.
Description	The name of the solution and user inputted comments
# Scan	The number of the scan.
Scan From	Select Scan from
Scans To	Select Scan To
Max OD	The OD cutoff of the absorbance.
Cell/Channel	Cell and channel column
Lambda	The wavelength(s) samples were measured by.
Reference	Define the reference scans in this column.
Save	Select which Cell/Channel to save.
Previous	Navigate to the previous wavelength.
Next	Navigate to the next wavelength.
Lambda Start	Start range of the wavelengths detected.
Lambda End	End range of the wavelengths detected.
Shift to Zero	Select region at baseline and shift whole scan to baseline.
Pick Region	Define the region to baseline.
Default	Use default parameters.
Save Absorbance Data	Save the absorbance data.
Status	Status bar.
Plot Both Intensity and Reference Scans	checkbox to include both sample and reference scans.
Intensity Plot	Plot of Intensity data, before subtraction.
Absorbance Plot	Plot of absorbance data, after subtraction.
Help	Open help documentation of this module.
Close	Close window.