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Main Authors: Kalinin, Alexandr A., Llanos, Paula, Sommer, Theresa Maria, Sestini, Giovanni, Hou, Xinhai, Sexton, Jonathan Z., Wan, Xiang, Dinov, Ivo D., Athey, Brian D., Rivron, Nicolas, Carpenter, Anne E., Cimini, Beth, Singh, Shantanu, O'Meara, Matthew J.
Format: Preprint
Published: 2025
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Online Access:https://arxiv.org/abs/2507.05383
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author Kalinin, Alexandr A.
Llanos, Paula
Sommer, Theresa Maria
Sestini, Giovanni
Hou, Xinhai
Sexton, Jonathan Z.
Wan, Xiang
Dinov, Ivo D.
Athey, Brian D.
Rivron, Nicolas
Carpenter, Anne E.
Cimini, Beth
Singh, Shantanu
O'Meara, Matthew J.
author_facet Kalinin, Alexandr A.
Llanos, Paula
Sommer, Theresa Maria
Sestini, Giovanni
Hou, Xinhai
Sexton, Jonathan Z.
Wan, Xiang
Dinov, Ivo D.
Athey, Brian D.
Rivron, Nicolas
Carpenter, Anne E.
Cimini, Beth
Singh, Shantanu
O'Meara, Matthew J.
contents Microscopy enables direct observation of cellular morphology in 3D, with transmitted-light methods offering low-cost, minimally invasive imaging and fluorescence microscopy providing specificity and contrast. Virtual staining combines these strengths by using machine learning to predict fluorescence images from label-free inputs. However, training of existing methods typically relies on loss functions that treat all pixels equally, thus reproducing background noise and artifacts instead of focusing on biologically meaningful signals. We introduce Spotlight, a simple yet powerful virtual staining approach that guides the model to focus on relevant cellular structures. Spotlight uses histogram-based foreground estimation to mask pixel-wise loss and to calculate a Dice loss on soft-thresholded predictions for shape-aware learning. Applied to a 3D benchmark dataset, Spotlight improves morphological representation while preserving pixel-level accuracy, resulting in virtual stains better suited for downstream tasks such as segmentation and profiling.
format Preprint
id arxiv_https___arxiv_org_abs_2507_05383
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Foreground-aware Virtual Staining for Accurate 3D Cell Morphological Profiling
Kalinin, Alexandr A.
Llanos, Paula
Sommer, Theresa Maria
Sestini, Giovanni
Hou, Xinhai
Sexton, Jonathan Z.
Wan, Xiang
Dinov, Ivo D.
Athey, Brian D.
Rivron, Nicolas
Carpenter, Anne E.
Cimini, Beth
Singh, Shantanu
O'Meara, Matthew J.
Computer Vision and Pattern Recognition
Quantitative Methods
I.4.9; J.3
Microscopy enables direct observation of cellular morphology in 3D, with transmitted-light methods offering low-cost, minimally invasive imaging and fluorescence microscopy providing specificity and contrast. Virtual staining combines these strengths by using machine learning to predict fluorescence images from label-free inputs. However, training of existing methods typically relies on loss functions that treat all pixels equally, thus reproducing background noise and artifacts instead of focusing on biologically meaningful signals. We introduce Spotlight, a simple yet powerful virtual staining approach that guides the model to focus on relevant cellular structures. Spotlight uses histogram-based foreground estimation to mask pixel-wise loss and to calculate a Dice loss on soft-thresholded predictions for shape-aware learning. Applied to a 3D benchmark dataset, Spotlight improves morphological representation while preserving pixel-level accuracy, resulting in virtual stains better suited for downstream tasks such as segmentation and profiling.
title Foreground-aware Virtual Staining for Accurate 3D Cell Morphological Profiling
topic Computer Vision and Pattern Recognition
Quantitative Methods
I.4.9; J.3
url https://arxiv.org/abs/2507.05383