Multi‐Photon Microscopy

Sanderson, Jeremy

doi:10.1002/cpz1.634

Cited by 6 publications

(3 citation statements)

References 160 publications

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“…While we demonstrate that this confocal-based method can achieve stable longterm imaging deep into intact tissue, it should be noted that multiphoton microscopes cause less photodamage and penetrate to greater depths compared with confocal instruments 17 . Therefore, the utility of this tool will rely on the strength of the fluorescent signal in the transgenic animal model of choice as well as the tissue depth required to achieve experimental goals.…”

Section: Figure Legendsmentioning

confidence: 67%

Streamlined intravital imaging approach for long-term monitoring of epithelial cell dynamics on an inverted confocal microscope

Hamersky,

Tekale,

Winfree

et al. 2023

Preprint

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Understanding normal and aberrant in vivo cell behaviors is necessary to develop clinical interventions to thwart disease initiation and progression. It is therefore critical to optimize imaging approaches that facilitate the observation of cell dynamics in situ, where tissue structure and composition remain unperturbed. The epidermis is the body's outermost barrier as well as the source of the most prevalent human cancers, namely cutaneous skin carcinomas. The accessibility of skin tissue presents a unique opportunity to monitor epithelial and dermal cell behaviors in intact animals using noninvasive intravital microscopy. Nevertheless, this sophisticated imaging approach has primarily been achieved using upright multiphoton microscopes, which represents a significant barrier-for-entry for most investigators. In this study, we present a custom-designed 3D-printed microscope stage insert suitable for use with inverted confocal microscopes that streamlines long-term intravital imaging of ear skin in live transgenic mice. We believe this versatile invention, which may be customized to fit the inverted microscope brand and model of choice, as well as adapted to image additional organ systems, will prove invaluable to the greater scientific research community by significantly enhancing the accessibility of intravital microscopy. This technological advancement is critical to bolster our understanding of live cell dynamics in both normal and disease contexts.

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Section: Figure Legendsmentioning

confidence: 67%

Streamlined intravital imaging approach for long-term monitoring of epithelial cell dynamics on an inverted confocal microscope

Hamersky,

Tekale,

Winfree

et al. 2023

Preprint

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“…Regardless of the magnification and numerical aperture of the objective lens used, the imaging throughput of current microscopes is typically only in the order of ten megapixels [ 1 ]. This leads to a compromise between high resolution and Field of View (FOV) when imaging.…”

Section: Introductionmentioning

confidence: 99%

A Residual Dense Attention Generative Adversarial Network for Microscopic Image Super-Resolution

Liu,

Weng,

Gao

et al. 2024

Sensors

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With the development of deep learning, the Super-Resolution (SR) reconstruction of microscopic images has improved significantly. However, the scarcity of microscopic images for training, the underutilization of hierarchical features in original Low-Resolution (LR) images, and the high-frequency noise unrelated with the image structure generated during the reconstruction process are still challenges in the Single Image Super-Resolution (SISR) field. Faced with these issues, we first collected sufficient microscopic images through Motic, a company engaged in the design and production of optical and digital microscopes, to establish a dataset. Secondly, we proposed a Residual Dense Attention Generative Adversarial Network (RDAGAN). The network comprises a generator, an image discriminator, and a feature discriminator. The generator includes a Residual Dense Block (RDB) and a Convolutional Block Attention Module (CBAM), focusing on extracting the hierarchical features of the original LR image. Simultaneously, the added feature discriminator enables the network to generate high-frequency features pertinent to the image’s structure. Finally, we conducted experimental analysis and compared our model with six classic models. Compared with the best model, our model improved PSNR and SSIM by about 1.5 dB and 0.2, respectively.

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“…Label-free multiphoton microscopy allows for the simultaneous acquisition of both detailed structural and chemical information within biological samples. While commonly used in biomedical research for exciting and analyzing fluorescent dyes (Sanderson 2023 ), multiphoton microscopy can provide chemical selectivity by exploiting the simultaneous acquisition of multiple nonlinear signals originating directly from endogenous tissue constituents.…”

Section: Introductionmentioning

confidence: 99%

Vibrational spectroscopy and multiphoton microscopy for label-free visualization of nervous system degeneration and regeneration

Galli,

Uckermann

2023

Biophys Rev

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Neurological disorders, including spinal cord injury, peripheral nerve injury, traumatic brain injury, and neurodegenerative diseases, pose significant challenges in terms of diagnosis, treatment, and understanding the underlying pathophysiological processes. Label-free multiphoton microscopy techniques, such as coherent Raman scattering, two-photon excited autofluorescence, and second and third harmonic generation microscopy, have emerged as powerful tools for visualizing nervous tissue with high resolution and without the need for exogenous labels. Coherent Raman scattering processes as well as third harmonic generation enable label-free visualization of myelin sheaths, while their combination with two-photon excited autofluorescence and second harmonic generation allows for a more comprehensive tissue visualization. They have shown promise in assessing the efficacy of therapeutic interventions and may have future applications in clinical diagnostics. In addition to multiphoton microscopy, vibrational spectroscopy methods such as infrared and Raman spectroscopy offer insights into the molecular signatures of injured nervous tissues and hold potential as diagnostic markers. This review summarizes the application of these label-free optical techniques in preclinical models and illustrates their potential in the diagnosis and treatment of neurological disorders with a special focus on injury, degeneration, and regeneration. Furthermore, it addresses current advancements and challenges for bridging the gap between research findings and their practical applications in a clinical setting.

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Multi‐Photon Microscopy

Cited by 6 publications

References 160 publications

Streamlined intravital imaging approach for long-term monitoring of epithelial cell dynamics on an inverted confocal microscope

Streamlined intravital imaging approach for long-term monitoring of epithelial cell dynamics on an inverted confocal microscope

A Residual Dense Attention Generative Adversarial Network for Microscopic Image Super-Resolution

Vibrational spectroscopy and multiphoton microscopy for label-free visualization of nervous system degeneration and regeneration

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