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Investigation of Drug Responses in 3-D Tumor Spheroid Models Using Two-Photon Scanning Structured Illumination Super-Resolution Microscopy With Frequency-Specific Denoising Enhancement
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DOI：10.1109/tmi.2026.3698950.png)
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Abstract
En 中文
Two-dimensional cell culture models have long been a cornerstone of biomedical research; however, they often fail to accurately replicate the in vivo environment. In recent years, three-dimensional (3D) cell cultures, particularly 3D spheroid models, have gained recognition for their ability to better mimic the complexities of the in vivo environment, making them valuable tools for studying cellular behavior and responses. Tumor spheroids, in particular, have significant applications in anticancer therapy evaluation, providing a more physiologically relevant model by simulating the spatial architecture and microenvironment of tumors. However, due to the limitations imposed by optical diffraction and background noise in 3D imaging, traditional imaging methods are unable to accurately resolve the growth, morphological changes, and drug responses of tumor spheroids. To address this issue, super-resolution imaging technologies have emerged. Structured illumination microscopy (SIM) combined with reconstruction algorithms can effectively enhance resolution, but challenges such as limited light penetration of single-photon imaging and high background noise remain in 3D imaging. In this paper, an advanced SIM technology with large depth and low noise 3D imaging capability is developed. This study introduces a novel frequency-specific denoising method (FSDM) to effectively reduce noise through adjusting the weights of high-frequency signals to preserve image details. The FSDM optimization significantly reduces background interference from deeper tissue layers, improving image details and the overall quality of 3D imaging. For the first time, scanning SIM is integrated with two-photon microscopy (TPEF-SIM) for 3D imaging, leveraging the strengths of both techniques to enhance resolution and overcome light penetration limitations.
Keywords:
Structured illumination microscopy
two-photon
tumor spheroid models
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