Iterative Bleaching Extends multi-pleXity (IBEX) imaging method, mouse spleen

This dataset was acquired using the Iterative Bleaching Extends multi-pleXity (IBEX) imaging method described in: “IBEX: A versatile multi-plex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues“, A. Radtke et al., 2020, doi:10.1073/pnas.2018488117.

It is comprised of a three cycle IBEX experiment performed on mouse spleen sections labeled with the nuclear marker JOJO-1 and membrane label CD4 AF594. Images were acquired using an inverted Leica TCS SP8 X confocal microscope equipped with a 40X objective (NA 1.3), 4 HyD and 1 PMT detectors, a white light laser that produces a continuous spectral output between 470 and 670 nm as well as 405, 685, and 730 nm lasers. All images were captured at an 8-bit depth, with a line average of 3, and 1024x1024 format with the following pixel dimensions: x (0.284 mm), y (0.284 mm), and z (1 mm). Images were tiled and merged using the LAS X Navigator software (LAS X 3.5.5.19976).

Markers per channel in each of the three cycles:

1. spleen_panel1.nrrd (6 channels): B220 PE, CD8 BV421, IgD AF700, CD4 AF594, JOJO, Foxp3 eF660

2. spleen_panel2.nrrd (7 channels): CD169 PE, F480 BV421, MHCII AF700, CollIV AF488, JOJO, CD11c AF647, CD4 AF594

3. spleen_panel3.nrrd (7 channels): CD31 PE, CD68 BV421, Ki67 AF700, CD45 AF488, CD4 AF594, JOJO, CD3 AF647

The panels can be registered using the code available on github: https://github.com/niaid/sitk-ibex

To view these multi-channel images, in nrrd format, use the Fiji viewer. The data is stored in XYZC order.