Multichannel analytics development

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=March 2011=
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=April 2011=
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=July 2011=
  
 
=August 2011=
 
=August 2011=
  
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The Roysam lab focused on pipeline integration in order to reveal kinks in the planned workflow.
  
== Image Preprocessing Improvements ==
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== Image Preprocessing Additions ==
  
*Background subtraction used to remove the blur and imaging artifacts
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*A background subtraction step to remove the low-frequency noise and large imaging artifacts from the raw images. 
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*A normalization step to equalize foreground contrast and correct for changes in overall brightness as a function of depth.
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These additions allowed us to montage a few data sets which had previously defied automated registration.
  
 
== Tiling Functions for Microglia Tracing and Nuclei Segmentation: ==
 
== Tiling Functions for Microglia Tracing and Nuclei Segmentation: ==
  
 
*Montage can be split into smaller tiles for processing.
 
*Montage can be split into smaller tiles for processing.
*Workflow of individual tiles can be distributed.
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*Workflow of individual tiles can be distributed more easily:
**Tracing Tiles of Microglia
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**Tracing microglial tiles
**Segment and Classify Nuclei
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**Segmentation and classification of nuclei tiles
*Resulting Files can be stitched back into Global Coordinates
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*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 ==
 
== Soma Detection and Segmentation Improvements ==
  
*Old method of morphological functions on the GFP channel missed somas.
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*Bug: previous method of morphological functions on the microglia channel missed some somata.
*Correlate Nuclei Channel to Microglia channel to identify somas
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*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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=September 2011=
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== Stress Test - Data 9/12 ==
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*Largest to-date data set (~48 GB per channel) processed
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**Minor bug relating to data registration uncovered, but effort largely successful
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=October 2011=
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== Pipeline Improvements ==
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*Added a single-tile analysis step for quick verification of batch processing validity.

Latest revision as of 21:00, 11 October 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.

September 2011

Stress Test - Data 9/12

  • Largest to-date data set (~48 GB per channel) processed
    • Minor bug relating to data registration uncovered, but effort largely successful

October 2011

Pipeline Improvements

  • Added a single-tile analysis step for quick verification of batch processing validity.
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