Viability

Experimental protocol

Detailed procedures for preparing and recording images of filarial nematode microfilariae (mf) can be found at the following link: Bivariate, high-content screening of Brugia malayi microfilariae

Configuration of the GUI

Viability can be analyzed using a generic Segmentation pipeline directed to the proper wavelength (if using a fluorescent indicator). For microfilaria viability specifically, there is also a CellProfiler pipeline available.

Expected input

Viability data should be analyzed in the form of individual TIF images per frame (i.e., the TimePoint structure utilized by ImageXpress). See the Data Organization page for more details. The plate directory should have a single TimePoint with individual images for each well:

Viability file structure

All experiments should include a single wavelength. Multisite images should be stitched according the ImageXpress + Multi Site instructions. The above screenshot shows the structure of unstitched input images that include the _s[1|2|3|4].TIF structure.

Validated species and stages

Filarial nematodes (i.e., Brugia malayi and Dirofilaria immitis)

  • Microfiliariae
  • L3s
  • Adults (with varying success)

Caenorhabditis elegans

  • Larvae
  • Young adults
  • Gravid adults

Example plates

  • 20210917-p15-NJW_913: Brugia malayi microfilariae

Expected output

A CSV file with at least two columns or many columns of CellProfiler output, depending on the selected pipeline. If using Metadata, there will be an additional column for each provided metadata data frame.