Multiscale and multimodal image fusion. Coping with differences in scanned area and spatial resolution for Raman/fluorescence images of labelled cells.

06 February 2025, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Multiscale and multimodal image fusion is a challenge derived from the diversity of chemical and spatial information provided by the current hyperspectral image platforms. Efficient image fusion approaches are essential to exploit the complementary chemical information across different zoom scales. Most current image fusion algorithms tend to work by equalizing the spatial characteristics of the platforms to be combined, i.e., downsampling pixel size and cropping non-common scanned sample areas if required. In this work, a new image unmixing algorithm based on a flexible mathematical framework is proposed to enable working with all available image information while preserving the original spatial properties of every imaging measurement. The algorithm is tested on a challenging image fusion scenario of fluorescence and Raman images collected on labelled HeLa cells. The system is relevant from an analytical point of view, since smart fluorescence labelling allows profiting from the excellent morphological information without causing interferences in the rich chemical information furnished by Raman. From a data handling perspective, it offers a challenging multiscale problem, where the fast fluorescence imaging acquisition allows recording full cell images, and the slower Raman image acquisition is focused on scanning only relevant small regions of the cells analyzed. By applying the image fusion algorithm proposed, an improved morphological and chemical characterization of cell constituents in the full cell area is obtained despite the different spatial scales used in the original imaging measurements.

Keywords

Raman
Fluorescence
Image fusion
Fluorescence labelling
Image unmixing
Multiscale
Multimodal

Supplementary materials

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Title
Supporting information. Multiscale and multimodal image fusion. Coping with differences in scanned area and spatial resolution for Raman/fluorescence images of labelled cells
Description
The supporting information includes fluorescence spectral information of the labelling agents used in figures S1 and S2. Figures S3 and S4 show images and/or spectra before and after preprocessing in fluorescence and Raman images, respectively. A graphical explanation of the modified least squares steps in incomplete multiset MCR-ALS analysis is displayed in figure S5. Full results of fluorescence and Raman individual MCR-ALS analyses can be consulted in figures S6 and S7, respectively. Table S1 contains the Raman features used for component identification in Raman and image fusion analyses. Figure S8 displays all the results related to the fused images from the incomplete multiset analysis. Figure S9 shows the fluorescence distribution maps recovered from the incomplete multiset analysis.
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