Two-Photon Microscopy for Deep Tissue Imaging of Living Specimens

Franke, Tilman; Rhode, Sebastian

doi:10.1017/s1551929512000430

Cited by 4 publications

(4 citation statements)

References 9 publications

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“…Howerver the method has not been used yet for repetitive imaging. Barbosa et al, 2015a;Dray et al, 2015;Guesmi et al, 2018 activity. Genetic tracing and birth dating experiments indicate that most, likely all, RG of the dorso-medial and anterior pallial domain originate from a constitutively neurogenic lineage, i.e., generating neurons without interruption from embryo to adult (Figure 2A; Dirian et al, 2014;Furlan et al, 2017).…”

Section: Furlan Et Al 2017mentioning

confidence: 99%

Conserved and Divergent Features of Adult Neurogenesis in Zebrafish

Labusch

Mancini

Morizet

et al. 2020

Front. Cell Dev. Biol.

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Adult neurogenesis, i.e., the generation of neurons from neural stem cells (NSCs) in the adult brain, contributes to brain plasticity in all vertebrates. It varies, however, greatly in extent, location and physiological characteristics between species. During the last decade, the teleost zebrafish (D. rerio) was increasingly used to study the molecular and cellular properties of adult NSCs, in particular as a prominent NSC population was discovered at the ventricular surface of the dorsal telencephalon (pallium), in territories homologous to the adult neurogenic niches of rodents. This model, for its specific features (large NSC population, amenability to intravital imaging, high regenerative capacity) allowed rapid progress in the characterization of basic adult NSC features. We review here these findings, with specific comparisons with the situation in rodents. We specifically discuss the cellular nature of NSCs (astroglial or neuroepithelial cells), their heterogeneities and their neurogenic lineages, and the mechanisms controlling NSC quiescence and fate choices, which all impact the neurogenic output. We further discuss the regulation of NSC activity in response to physiological triggers and non-physiological conditions such as regenerative contexts.

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Section: Furlan Et Al 2017mentioning

confidence: 99%

Conserved and Divergent Features of Adult Neurogenesis in Zebrafish

Labusch

Mancini

Morizet

et al. 2020

Front. Cell Dev. Biol.

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“…Throughout the surgery and imaging, ophthalmic ointment was applied to avoid drying of the animal's eyes. To reach the deeper pancreas tissue, 2photon microscopy was utilized, with penetration depth reaching from 500 µm up to 1 mm into the tissue (58,59). Animals were imaged using a Leica TCS SP8 DIVE confocal multiphoton system mounted on an inverted stand equipped with a Spectra-Physics MaiTai DeepSee laser and mounted with a 40x 1.1 NA water objective.…”

Section: Intravital Imaging Of Endogenous Mouse Pancreasmentioning

confidence: 99%

Real TimeIn vivoAnalysis of Pancreatic Beta-cell Autophagic Flux Reveals Impairment Before Onset of Autoimmune Diabetes

Melnyk,

Muralidharan,

Duffett

et al. 2023

Preprint

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The catabolic pathway of autophagy is critical for pancreatic beta-cell function and is defective in established type 1 diabetes (T1D). However, it is unclear when and how this critical cell process becomes defective during diabetes pathogenesis. To study the nature of autophagy dysfunction in the context of autoimmune diabetes, we utilized intravital microscopy to study autophagic flux in vivo in real time. We generated a custom AAV8-packaged mCherry-eGFP-LC3B biosensor driven by the insulin promoter for beta-cell-selective expression. For real time autophagic flux evaluation, fluorescent signal from eGFP and mCherry fluorophores was correlated in space and time to follow the process of autophagosome-lysosome fusion. We observed autophagic flux defects in the beta-cells of non-obese diabetic (NOD) mouse model of T1D prior to hyperglycemia onset that were less apparent in mice without a functional immune system. We also evaluated autophagic flux in human donor islets that were transplanted under the kidney capsule of immune incompetent mice. Collectively, we provide the first evaluation of autophagic flux in vivo in 4D and demonstrate that autophagy defects precede hyperglycemia in NOD mice suggesting a potential causative role for these defects in beta-cell demise during T1D pathogenesis.

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“…In addition, the laser was scanned and images of the focal plane were acquired along the z-axis, thus allowing imaging of the entire tissue [ 29 ]. Multiphoton fluorescence microscopy is based on the basic principle of two-photon excitation by ultrashort laser pulses and overcomes the disadvantages of confocal fluorescence microscopy (low tissue penetration and high invasiveness) [ 30 , 31 ]; this multiphoton fluorescence microscope uses long-wavelength excitation light, which has high tissue penetration and is suitable for live imaging [ 30 , 31 ].…”

Section: Tissue Clearing and Imagingmentioning

confidence: 99%

Understanding Breast Cancers through Spatial and High-Resolution Visualization Using Imaging Technologies

Takahashi

Kawahara

Kikuchi

2022

Cancers

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Breast cancer is the most common cancer affecting women worldwide. Although many analyses and treatments have traditionally targeted the breast cancer cells themselves, recent studies have focused on investigating entire cancer tissues, including breast cancer cells. To understand the structure of breast cancer tissues, including breast cancer cells, it is necessary to investigate the three-dimensional location of the cells and/or proteins comprising the tissues and to clarify the relationship between the three-dimensional structure and malignant transformation or metastasis of breast cancers. In this review, we aim to summarize the methods for analyzing the three-dimensional structure of breast cancer tissue, paying particular attention to the recent technological advances in the combination of the tissue-clearing method and optical three-dimensional imaging. We also aimed to identify the latest methods for exploring the relationship between the three-dimensional cell arrangement in breast cancer tissues and the gene expression of each cell. Finally, we aimed to describe the three-dimensional imaging features of breast cancer tissues using noninvasive photoacoustic imaging methods.

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Two-Photon Microscopy for Deep Tissue Imaging of Living Specimens

Cited by 4 publications

References 9 publications

Conserved and Divergent Features of Adult Neurogenesis in Zebrafish

Conserved and Divergent Features of Adult Neurogenesis in Zebrafish

Real TimeIn vivoAnalysis of Pancreatic Beta-cell Autophagic Flux Reveals Impairment Before Onset of Autoimmune Diabetes

Understanding Breast Cancers through Spatial and High-Resolution Visualization Using Imaging Technologies

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