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Wave 2 Data: Live-cell time-lapse microscopy of pyroptosis.

This dataset contains live-cell time-lapse microscopy of pyroptosis in Acute Myloid Leukemia (AML) cells. The dataset was generated by Saguaro BioSciences and University of Montreal's imaging platform in collaboration with the lab of Guy Sauvageau, PhD.

Experimental Design

The dataset was generated by imaging AML cells that were treated with pyroptosis-inducing agents. The cells were imaged every ~11 minutes for 20 hours using a 20x objective on a Yokogawa CQ1 cell voyager spinning disk confocal microscope. The images were acquired with the following parameters:

  • 20x objective - NA 0.80
  • 56 wells on a 384-well plate
  • 4 fields of view (FOV) per well
  • 1 z-slice per FOV
  • 5 channels

The images were acquired in 5 channels:

Channel # Channel Name Excitation (nm) Emission (nm) Dye Exposure (ms)
1 ChromaLIVE 640 685/40 ChromaLIVE 50
2 ChromaLIVE 488 685/40 ChromaLIVE 200
3 SYTOX Green 488 525/50 SYTOX Green 50
4 Nuclei 405 447/60 NulceoLive Blue 50
5 Brightfield 50

The wave 2 plate map can be viewed in the Wave2_data/platemap_readme.md.

Data

This wave 2 data is preliminary and generated to identify the best conditions for imaging pyroptosis in AML cells. The data has the following naming convention:

  • {well}_{FOV}_{time}_{zslice}_{channel}.tif Where:
  • well is the well number W0001 is the top left well and W0384 is the bottom right well. The well numbers snake through the plate, so W0001 is the top left well, W0002 is the second well in the top row. W0025 is the first well in the second row, and so on.
  • FOV is the field of view number, with F0001 being the top left FOV and FOV4 being the bottom right FOV.
  • time is the timepoint in minutes since the start of the experiment, with T0001 being the first timepoint at 0 minutes. The timepoints are every X minutes, so T0002 is the second timepoint at X*2 minutes, and so on.
  • zslice is the z-slice number, with Z0001 being the first z-slice.
  • channel is the channel number, with C1 being the first channel

Processing

Preprocessing

These raw data are preprocessed into single well, single field of view, single timepoint, single z-slice images directories with all channels. This allows for each FOV to be processed independently.

Illumination Correction

Image-based profiling

Image-based profile processing