Deconvolution, Visualization and Analysis Software for Widefield and Confocal Microscopy
Media Cybernetics is committed to delivering the most advanced algorithms available on the
market today by the cutting edge research to develop the most advanced algorithms for
tomorrow.Media Cybernetics provides uncompromised
results with faster performance than ever before.
The best just keeps getting better and faster…
Why Deconvolution ?
Each single plane contains blur from planes above and below
PSF: 3D representation you get when imaging a sub-resolution latex bead
Diffraction and aberration in the optical path lead to Blur / Haze
3-D diffraction pattern (= Point Spread Function PSF)
Blurred, faint
Sharp, bright
Blurred, faint
z
What is “Deconvolution” ?
“Deconvolution” is a computational procedure
that restores the quality of images.
Usage:
• Microscopy: biomedical research, ophthalmology, medical diagnostics, nuclear imaging (CT), food processing, forensics, skin care/beauty product development, etc.
• Astronomy: telescope imaging, satellite imaging, aerial imaging (cartography), etc.
• Industrial: Quality control for printed circuit boards and semiconductors, etc.
(Spirogyra)
Benefits of Deconvolution
1. Haze removal 1. Haze removal More contrast More contrast Better segmentation Better segmentation
2. Improved resolution 2. Improved resolution More detail revealed More detail revealed
3. Access to depth information3. Access to depth information
4. Produces quantitative results (non-destructive)4. Produces quantitative results (non-destructive)
Deconvolution vs. Deblurring
• Deconvolution– More closely approximates a
reversal of the blurring process
– Produces Quantitative results (reverses blurring accurately)
– Better representation of actual morphology
• Deblurring– Can make images look better
fast.– Results are strictly qualitative – Produces artifacts– Morphologically unreliable.
Qualitative vs. Quantitative
No-Neighbors Single slice haze removal.
Nearest-Neighbors Haze removal from a few optical sections.
Inverse filtering Inversion of the haze using a theoretical PSF.
Constrained iterative Iterative deconvolution with a fixed PSF.
Maximum likelihood Max likelihood estimate-based iterativedeconvolution with a fixed PSF.
Adaptive blind Max likelihood estimate-based iterativedeconvolution deconvolution deriving the PSF from
information found in the ‘haze’ itself.
No/Nearest Neighbors
• Uses center slice and two adjacent slices• Estimate amount of blur that adjacent slices contribute to center
slice• Subtract away the blur (remove photons)• Deblur one slice at a time• Fast, 2D, only qualitative (unsharp mask)
0
+z
-z
Inverse/Wiener Filter
• Restoration algorithm – looks at whole structure• Works well on large objects• Fast, 3D, qualitative• Regularization/Stability
ImageObject PSF
DeblurredImagePSF
1
Imaging:
Inverse Filter:
Constrained Iterative Deconvolution
• Requires PSF (theoretical or measured)• Iteratively improve object estimate • Impose constraints (non-negativity)• Requires highly optimized imaging system
ReblurredObjectEstimate
PSF Original Blurred
ImprovedEstimate
-
Update Error
Iterative Blind Deconvolution
• Estimate both PSF and Object!• Need microscope parameters (NA, RI, xyz, λ)• Theoretical PSF with spatial/spectral constraints• Object with non-negativity and noise constraints
ObjectEstimate Update
ObjectPSF
Estimate
UpdatePSF
OriginalBlurred
Why AQI and “Blind” Deconvolution ?
• Ease of Use– No need to image beads and measure PSF
• Much less expertise required by the user• Less time required on the microscope• Allows for processing of existing data• Runs on PC• Allows for deconvolution of large images• The reconstructed PSF can vary over the volume• Disadvantages
– More processing time
When to use what?
When Speed & Haze removal are important
No Neighbor
Nearest Neighbor
Inverse Filter
3D Deconvolution
When Quality & Quantitation are critical
XY Max Projections
XY Max Projections
XZ Max Projections
XZ Max Projections
Raw
InverseFilter
Nearest Neighbors
Blind Deconvolution
2D Blind Deconvolution – true iterative blind deconvolution
• No image parameters required• Ideal for optically thin specimens
• Assumes everything is in one plane of focus
Why Deconvolve?
• Improved resolution of small features
• Better definition of structure in multiple dimensions
• More accurate localization of intensity for quantification and ratiometry
• Signal to noise (SNR) improvement
Why Media Cybernetics…
• Established technological focus in deconvolution – over 25
years of dedicated research
• Breadth of algorithms that are well known and accepted world-
wide – optimized for maximum robustness
• Media Cybernetics stands behind all of its products and
provides world class education and support.
• Committed focus to deconvolution, visualization and analysis
R&D ensures customer access to the latest technologies for
today and tomorrow.
Media Cybernetics:Offering complete post-processing solutions
• Image improvement, restoration, visualization and analysis that facilitates more accurate and compelling results.
• Media Cybernetics creates the tools to help you do the science.
• AQI Product Line– AutoDeblur (2D, Silver, Gold)– AutoVisualize (with improved 3D-Viewer)– AutoAnalyze (FRET, Ratio, Object Counting/Tracking,
Colocalization)– Tools (automatic, manual and channel-channel alignment)
• Image-Pro Product Line– Image-Pro Plus and Image-Pro Plug-In module bundles– Express– Analyzer