The sources of image degradation can be divided into four independent phenomena: noise, scatter, glare, and blur. While all of these apply for widefield microscopy, application of a pinhole eliminates almost all sources of degradation except noise.

Deconvolution is a computationally intensive image processing technique that is being increasingly utilized for improving the contrast and resolution of digital images captured in the microscope. It is capable of:

• Increasing resolution, especially in the axial direction
• Removing noise
• Increasing contrast

A few packages are available that perform deconvolution. Huygens from SVI (Scientific Volume Imaging) is the most popular one. A number of algorithms allows to address images acquired at different conditions and delivers superior quality. It is especially useful for the groups interested in visualizing small structures - microtubules, spines, receptors, etc.

Object analyzer tool for 3D microscopy images in Huygens Professional allows you to obtain statistics of individual objects by clicking on them, or analyzing all objects with a single button press.

The image is segmented into objects by an effective seed-threshold level, and connection technique. A third method that can be applied is the watershed method which increases the reliability of the segmentation further. To remove too small objects in an early stage from the analysis a garbage level can be set below which objects are discarded. After that, detected objects are automatically labeled and sent to a continuous Iso Surface Ray Tracing renderer.

Predefined filters allow to remove objects with different properties before further analysis.

The segmented and labeled image is shown as a colored iso-surface image. Each object is assigned a different color. Because the iso-surface renderer can recompute a new iso surface rapidly, the threshold can be manipulated interactively.

When and Where

Oct. 20, 2012  - Nov. 8, 2012

Rm. 8122, Data Analysis Station

How to Apply

Please contact Vladimir Ghukasyan to reserve time and get a training/help with deconvolution/object analysis.

Rm. 7109F NRB

Tel.: 919 966 5807

vladimir@email.unc.edu

Additional Information:

Huygens Deconvolution