Multichannel analytics development

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Revision as of 16:56, 6 September 2011

Contents

March 2011

April 2011

July 2011

August 2011

The Roysam lab focused on pipeline integration in order to reveal kinks in the planned workflow.

Image Preprocessing Additions

  • A background subtraction step to remove the low-frequency noise and large imaging artifacts from the raw images.
  • A normalization step to equalize foreground contrast and correct for changes in overall brightness as a function of depth.

These additions allowed us to montage a few data sets which had previously defied automated registration.

Tiling Functions for Microglia Tracing and Nuclei Segmentation:

  • Montage can be split into smaller tiles for processing.
  • Workflow of individual tiles can be distributed more easily:
    • Tracing microglial tiles
    • Segmentation and classification of nuclei tiles
  • Resulting files can be stitched back into global coordinates automatically, since the output of each tile only needs a static translation.

Soma Detection and Segmentation Improvements

  • Bug: previous method of morphological functions on the microglia channel missed some somata.
  • Fix: combining the DAPI channel with the microglia channel reduced extrasomatic noise, allowing us to use more relaxed parameters to identify somata.
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