Imaging of biological specimens brings a wealth of information about cell structure and function; as such, it is one of the most used techniques in biological research. Proper analysis of image data requires high quality images. Deconvolution is one process that ensures high quality data as it removes the haze and blur and restores important details to datasets.
AutoDeblur offers the most powerful deconvolution tools and is the leading image deconvolution software. It allows the user to retrieve better data from images using 2D and 3D algorithms. The Blind Deconvolution algorithm is unique in the industry and unlike other deconvolution products, AutoDeblur does not require the manual calibration and measurement of the point spread function which is calculated directly from the collected data set. AutoDeblur is very versatile and can be used with widefield epi-fluorescence, DIC, transmitted-light bright field, two-photon and spinning disk confocal microscopes.
In addition to the full complement of algorithms, AutoDeblur has a multi-dimensional platform for 3D time-series datasets. Individual data sets can be processed as a single dataset and one can start working on another dataset once processing has begun. AutoDeblur allows the user to load an unlimited number of channels and one can customize the color wavelength, dye, etc. Multiple concurrent 5D viewers can be opened and synchronized for data comparisons. There is a multitude of algorithms one can use: 3D-blind or non-blind deconvolution, inverse filter, no/nearest neighbor, etc. There is also a spherical aberration detection and correction tool, as well as correction for photobleaching, optical density variability and other common imaging problems. Even multi-channel and time-series datasets are easily loaded and the program can calculate image spacing for time-saving and accurate image analysis.
Using 3D visualization and manipulation tools one can view time-series datasets through time as well as from multiple angles. The program also offers simultaneous viewing before and after 3D time-lapse volumes and the capability to scroll through slices of multiple datasets simultaneously and to create .AVI files of 3D data rotating through time. One can change channel color or brightness/contrast in real time.
Other features of the software include the Colocalization, Counting and Tracking modules. Easy to use ROI’s and threshold controls are used to obtain statistics about relative changes or concentration within datasets. In addition, image alignment, image smoothing, bleedthrough removal, background subtraction, and noise removal are available for more accurate results. It is also possible to work with time-series data to perform ratio imaging over time.
The program can also be used in the ratiometrics FRET module. It enables the researcher to see the graphic depiction of colocalized areas and to work with both single slice and full volume depictions of colocalized areas. One can also remove noise and improve object tracking accuracy, process and analyze multi-channel and 3D time-series datasets, export statistics into .csv format for further analysis, or create .AVI movies showing object motion. This module contains an impressive statistics feature and colocalization preview capabilities.
I have used AutoDeblur software from Media Cibernetics Inc. for 3D deconvolution of multiple channel and multiple timepoint data series obtained with a spinning disk confocal microscope of live Drosophila embryos injected with various rhodamine-labeled proteins and expressing various GFP-tagged proteins as well as fixed samples stained with various fluorochromes.
Postdoc
Department of Medical Genetics and Microbiology
University of Toronto