Dual deconvolution multifocal fluorescence response matrix microscopy for aberration-free high-throughput super-resolution imaging
Abstract
Fluorescence response matrix imaging enables separate computational correction of excitation and emission aberrations through dual deconvolution, but single-focus acquisition requires a large number of camera frames, limiting its practical use. Multifocal excitation can accelerate acquisition by recording many illumination positions in parallel, but denser foci also increase inter-focus crosstalk, which can bias the estimated excitation and emission optical transfer functions and degrade aberration correction. Here, we introduce high-throughput aberration-free fluorescence response matrix imaging with adaptive multifocal illumination, in which the inter-focus spacing of the multifocal lattice is adjusted according to specimen-dependent crosstalk. A programmable digital micromirror device generates multifocal excitation lattices, and the resulting detector-resolved fluorescence responses are assembled into a two-coordinate fluorescence response matrix. Dual deconvolution separately estimates and corrects the excitation and emission optical transfer functions, and the aberration-corrected matrix is then combined in the spatial-frequency domain to reconstruct the aberration-free image. This strategy reduces the number of required camera frames by approximately three orders of magnitude, allowing response matrices with ∼ 106 illumination positions to be acquired within seconds to a few minutes. The accelerated acquisition preserves the aberration-correction capability of dual deconvolution and achieves an estimated lateral resolution of ∼100 nm. Applied to heterogeneous mouse brain and kidney tissue, the method enables high-resolution, depth-resolved volumetric imaging while correcting specimen-dependent aberrations. Together, these results establish adaptive multifocal illumination as a practical route to high-throughput aberration-free fluorescence response matrix imaging.
Related
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BibTeX
@misc{lim2026dual,
title = {{Dual deconvolution multifocal fluorescence response matrix microscopy for aberration-free high-throughput super-resolution imaging}},
author = {Sumin Lim and Sanghyeon Park and Tran Thinh Le and Jin Hee Hong and Seokchan Yoon and Wonshik Choi},
howpublished = {arXiv},
year = {2026},
month = oct,
doi = {10.48550/arxiv.2610.15264},
eprint = {2610.15264},
archivePrefix = {arXiv},
url = {https://arxiv.org/abs/2610.15264},
}
RIS
TY - UNPB
TI - Dual deconvolution multifocal fluorescence response matrix microscopy for aberration-free high-throughput super-resolution imaging
AU - Sumin Lim
AU - Sanghyeon Park
AU - Tran Thinh Le
AU - Jin Hee Hong
AU - Seokchan Yoon
AU - Wonshik Choi
PY - 2026
DA - 2026/10/11
PB - arXiv
DO - 10.48550/arxiv.2610.15264
UR - https://arxiv.org/abs/2610.15264
AB - Fluorescence response matrix imaging enables separate computational correction of excitation and emission aberrations through dual deconvolution, but single-focus acquisition requires a large number of camera frames, limiting its practical use. Multifocal excitation can accelerate acquisition by recording many illumination positions in parallel, but denser foci also increase inter-focus crosstalk, which can bias the estimated excitation and emission optical transfer functions and degrade aberration correction. Here, we introduce high-throughput aberration-free fluorescence response matrix imaging with adaptive multifocal illumination, in which the inter-focus spacing of the multifocal lattice is adjusted according to specimen-dependent crosstalk. A programmable digital micromirror device generates multifocal excitation lattices, and the resulting detector-resolved fluorescence responses are assembled into a two-coordinate fluorescence response matrix. Dual deconvolution separately estimates and corrects the excitation and emission optical transfer functions, and the aberration-corrected matrix is then combined in the spatial-frequency domain to reconstruct the aberration-free image. This strategy reduces the number of required camera frames by approximately three orders of magnitude, allowing response matrices with ∼ 10^6 illumination positions to be acquired within seconds to a few minutes. The accelerated acquisition preserves the aberration-correction capability of dual deconvolution and achieves an estimated lateral resolution of ∼100 nm. Applied to heterogeneous mouse brain and kidney tissue, the method enables high-resolution, depth-resolved volumetric imaging while correcting specimen-dependent aberrations. Together, these results establish adaptive multifocal illumination as a practical route to high-throughput aberration-free fluorescence response matrix imaging.
ER -