ABSTRACT
Newly developed technologies have made it feasible to routinely collect highly multiplexed (20-60 channel) images at subcellular resolution from human tissues for research and diagnostic purposes. Extracting single cell data from such images requires efficient and accurate image segmentation, a challenging problem that has recently benefited from the use of deep learning. In this paper, we demonstrate two approaches to improving tissue segmentation that are applicable to multiple deep learning frameworks. The first uses “real augmentations” that comprise defocused and saturated image data collected on the same instruments as the actual data; using real augmentation improves model accuracy to a significantly greater degree than computational augmentation (Gaussian blurring). The second involves imaging the nuclear envelope to better identify nuclear outlines. The two approaches cumulatively and substantially improve segmentation on a wide range of tissue types and provide a set of improved segmentation models. We speculate that the use of real augmentations may have applications in image processing outside of microscopy.
Competing Interest Statement
PKS is a member of the SAB or BOD member of Applied Biomath, RareCyte Inc., and Glencoe Software, which distributes a commercial version of the OMERO database; PKS is also a member of the NanoString SAB. In the last five years the Sorger lab has received research funding from Novartis and Merck. Sorger declares that none of these relationships have influenced the content of this manuscript. SS is a consultant for RareCyte Inc. The other authors declare no outside interests.
Footnotes
Clarence Yapp clarence{at}hms.harvard.edu, Edward Novikov edward_novikov{at}seas.harvard.edu, Won-Dong Jang wdjang{at}seas.harvard.edu, Tuulia Vallius tuulia_vallius{at}hms.harvard.edu, Yu-An Chen yu-an_chen{at}hms.harvard.edu, Marcelo Cicconet marcelo_cicconet{at}hms.harvard.edu, Zoltan Maliga zoltan_maliga{at}hms.harvard.edu, Connor A. Jacobson connor_jacobson{at}hms.harvard.edu, Donglai Wei donglai{at}seas.harvard.edu, Sandro Santagata ssantagata{at}bics.bwh.harvard.edu, Hanspeter Pfister pfister{at}seas.harvard.edu, Peter K. Sorger peter_sorger{at}hms.harvard.edu
-Additional segmentation metrics added -Additional author added -Interobserver agreement metric added -Supplementary file updated