CVMar 29, 2025

scSplit: Bringing Severity Cognizance to Image Decomposition in Fluorescence Microscopy

arXiv:2503.22983v31 citationsh-index: 2Has Code
Originality Incremental advance
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This work addresses a specific limitation in computational multiplexing for life sciences by making image decomposition methods cognizant of varying intensity ratios, though it appears incremental as it builds on an existing iterative method.

The authors tackled the problem of image decomposition in fluorescence microscopy by proposing scSplit, a method that is aware of the unknown mixing ratios in superimposed images, and demonstrated its applicability on 5 public datasets for tasks like image splitting and bleedthrough removal.

Fluorescence microscopy, while being a key driver for progress in the life sciences, is also subject to technical limitations. To overcome them, computational multiplexing techniques have recently been proposed, which allow multiple cellular structures to be captured in a single image and later be unmixed. Existing image decomposition methods are trained on a set of superimposed input images and the respective unmixed target images. It is critical to note that the relative strength (mixing ratio) of the superimposed images for a given input is a priori unknown. However, existing methods are trained on a fixed intensity ratio of superimposed inputs, making them not cognizant of the range of relative intensities that can occur in fluorescence microscopy. In this work, we propose a novel method called scSplit that is cognizant of the severity of the above-mentioned mixing ratio. Our idea is based on InDI , a popular iterative method for image restoration, and an ideal starting point to embrace the unknown mixing ratio in any given input. We introduce (i) a suitably trained regressor network that predicts the degradation level (mixing ratio) of a given input image and (ii) a degradation-specific normalization module, enabling degradation-aware inference across all mixing ratios. We show that this method solves two relevant tasks in fluorescence microscopy, namely image splitting and bleedthrough removal, and empirically demonstrate the applicability of scSplit on 5 public datasets. The source code with pre-trained models is hosted at https://github.com/juglab/scSplit/.

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