DOI: 10.5061/dryad.gmsbcc2sq
title: Human intestine processed CODEX multiplexed images for donors B009-B012 (Part 2/2)

description: We performed CODEX (co-detection by indexing) multiplexed imaging on 64
 sections of the human intestine (~16 mm2) from 8 donors (B004, B005, B006,
 B008, B009, B010, B011, and B012) using a panel of 57
 oligonucleotide-barcoded antibodies. Subsequently, images underwent
 standard CODEX image processing (tile stitching, drift compensation, cycle
 concatenation, background subtraction, deconvolution, and determination of
 best focal plane). This data could be used to understand the cellular
 interactions, composition, and structure of the human intestine from the
 duodenum to the sigmoid colon and understand differences between different
 areas of the intestine. This data could be used as a healthy baseline to
 compare other single-cell datasets of the human intestine, particularly
 multiplexed imaging ones. The overall structure of the datasets is
 organized into 2 parts. This is the second part associated with this
 dataset are from donors B009, B010, B011, B012. This data is complimentary
 and can be linked to the single-cell processed dataframe already provided
 in another Dryad repository based on the donor name, folder name, and
 region number: https://doi.org/10.5061/dryad.pk0p2ngrf The raw imaging
 data can be found at (https://portal.hubmapconsortium.org/). We have
 created a landing page with links to all the raw dataset IDs and the
 HuBMAP ID for this Collection is HBM692.JRZB.356 and the DOI
 is:10.35079/HBM692.JRZB.356. This can be used to also pair it with the
 matched snRNAseq and snATACseq for each section of tissue.
description: For a detailed description of each of the steps of protocols and
 processes to obtain this data see the detailed materials and methods in
 the associated manuscript. Briefly, intestine pieces from 8 different
 sites across the small intestine and colon were taken and frozen in OCT.
 These were assembled into an array of 4 tissues, cut into 7 um slices, and
 stained with a panel of 57 CODEX DNA-oligonucleotide barcoded antibodies.
 Tissues were imaged with a Keyence microscope at 20x objective and then
 processed using image stitching, drift compensation, deconvolution, and
 cycle concatenation. 
description: The overall structure of the datasets is organized into 2 parts.
 This second part associated with this dataset is from donors B009, B010,
 B011, and B012. Each donor has a total of 8 tissue imaging regions of the
 different areas of the intestine (except for B009, which includes 1 extra
 folder of 4 images that were used as "test" imaging data). Each
 donor was split into two folders of 4 regions each stained and imaged at
 the same time in a 4-tissue array. For B004
 through B008, all small bowel (SB) or colon (CL) from the same donor were
 imaged together in the same array (hence the SB or CL designation in
 folder structure). For B009 through B012, each donor had 2 tissue regions
 from the small bowel and 2 tissue regions from the colon imaged together
 from the same array (hence the A or B designation in folder structure).
 These folder structure names will be useful to map back to the single cell
 CODEX dataframe (https://doi.org/10.5061/dryad.pk0p2ngrf) that connects each region back to its exact location within the small intestine or colon for each donor.  One region was imaged per tissue region per donor for a 8 total image stacks per donor in 2 different zipped folders (with the exception of B008_SB which has a duplicate imaged area of one region because this tissue was large enough on the coverslip to allow imaging of two separate areas). These image stacks have been zipped with the channelnames.txt file which describes the order of the fluorescent image stacks. The structure of this file is such that the markers are first ordered by the number of cycles, and then ordered within cycles in the following channel order: DAPI, then alexa488, then Cy3, then Cy5. Thus this provides the labels for the image stacks that are provided as a stacked TIF file that has one layer as the channels and the other layer as the cycles.  Most often we use FIJI (ImageJ) for visual analysis of the images and further image processing. However, QuPath and Python also offer functionality for opening large stacked tiff files.