Multichannel analytics development
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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.
