Two-photon microscopy of fungal keratitis-affected rabbit cornea ex vivo using moxifloxacin as a labeling agent

Lee, Jun Ho; Le, Viet-Hoan; Lee, Seunghun; Park, Jin Hyoung; Lee, Jin Ah; Tchah, Hungwon; Kim, Sungjee; Kim, Myoung Joon; Kim, Ki Hean

doi:10.1016/j.exer.2018.05.018

Cited by 9 publications

(8 citation statements)

References 23 publications

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“…Fine cellular details of animal-fungal pathogen interactions can also be visualized and studied by two-photon microscopy on thick viable lung slices (Hasenberg et al 2011 ) or in intact lungs (Bruns et al 2010 ) of mice infected by A. fumigatus. Using two-photon second harmonic generation microscopy, Lee et al ( 2018 ) were able to detect C. albicans and A. fumigatus cells in infected corneas in a fungal keratitis rabbit model ex vivo.…”

Section: Live-cell Imaging Time-lapse High-resolution and Special Mic...mentioning

confidence: 99%

“…The high translucency of host tissues towards red, far-red, and infrared light in animal-fungus and plant-fungus interaction studies has also fuelled the development of new long-wavelength reporting protein systems in a number of fungal species (Leal Jr. et al 2010 ; Su et al 2018 ; Vallarino et al 2018 ). In addition, red, far-red, and infrared light have been used successfully to study host-fungus interactions deep in tissues (Bruns et al 2010 ; Hasenberg et al 2011 ; Sørensen et al 2012 ; Vasilchenko et al 2016 ; Lee et al 2018 ). Combining red-shifted fluorophores with two-photon microscopy afforded increased spatial resolution in addition to better penetration (Tolić-Nørrelykke et al 2004 ; Lemar et al 2005 , 2007 ; García-Marcos et al 2008 ; Váchová et al 2009 ).…”

Section: Introductionmentioning

confidence: 99%

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Use of red, far-red, and near-infrared light in imaging of yeasts and filamentous fungi

Pócsi

Szigeti

Emri

et al. 2022

Appl Microbiol Biotechnol

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While phototoxicity can be a useful therapeutic modality not only for eliminating malignant cells but also in treating fungal infections, mycologists aiming to observe morphological changes or molecular events in fungi, especially when long observation periods or high light fluxes are warranted, encounter problems owed to altered regulatory pathways or even cell death caused by various photosensing mechanisms. Consequently, the ever expanding repertoire of visible fluorescent protein toolboxes and high-resolution microscopy methods designed to investigate fungi in vitro and in vivo need to comply with an additional requirement: to decrease the unwanted side effects of illumination. In addition to optimizing exposure, an obvious solution is red-shifted illumination, which, however, does not come without compromises. This review summarizes the interactions of fungi with light and the various molecular biology and technology approaches developed for exploring their functions on the molecular, cellular, and in vivo microscopic levels, and outlines the progress towards reducing phototoxicity through applying far-red and near-infrared light. Key points • Fungal biological processes alter upon illumination, also under the microscope • Red shifted fluorescent protein toolboxes decrease interference by illumination • Innovations like two-photon, lightsheet, and near IR microscopy reduce phototoxicity

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Section: Live-cell Imaging Time-lapse High-resolution and Special Mic...mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Use of red, far-red, and near-infrared light in imaging of yeasts and filamentous fungi

Pócsi

Szigeti

Emri

et al. 2022

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Two-photon microscopy [67] allowed study of host–pathogen interactions in real time and in three dimensions, with high resolution imaging up to a depth of 1 mm [68]. However, in the original use of this technique, mice had to be anaesthetised and the chest cavity opened to permit imaging of fungal infections, such as Aspergillus fumigatus [69,70]. Development of two-photon and multi-photon intravital microscopy [71,72,73], has provided insights into fungal trafficking and host–fungal interactions.…”

Section: Near-infrared Fluorescent Proteinmentioning

confidence: 99%

A Bright Future for Fluorescence Imaging of Fungi in Living Hosts

Chapuis

Ballou

MacCallum

2019

JoF

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Traditional in vivo investigation of fungal infection and new antifungal therapies in mouse models is usually carried out using post mortem methodologies. However, biomedical imaging techniques focusing on non-invasive techniques using bioluminescent and fluorescent proteins have become valuable tools. These new techniques address ethical concerns as they allow reduction in the number of animals required to evaluate new antifungal therapies. They also allow better understanding of the growth and spread of the pathogen during infection. In this review, we concentrate on imaging technologies using different fungal reporter proteins. We discuss the advantages and limitations of these different reporters and compare the efficacy of bioluminescent and fluorescent proteins for fungal research.

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“…Moxifloxacin, which is a fourth-generation fluoroquinolone, has been used as a cell-labeling agent in the imaging of various organs such as the eye, skin, bladder, brain, and colon of animal models. [12][13][14][15][16] Moxifloxacin was topically applied just like proflavine and labeled cell cytoplasm with intrinsic fluorescence. In the case of colon, goblet cells in the colonic crypt were visualized with strong fluorescence.…”

Section: Introductionmentioning

confidence: 99%

Feasibility of moxifloxacin and proflavine dual fluorescence imaging for detecting gastrointestinal neoplastic lesions: A prospective study

et al. 2023

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Objectives High‐contrast and high‐resolution imaging techniques would enable real‐time sensitive detection of the gastrointestinal lesions. This study aimed to investigate the feasibility of novel dual fluorescence imaging using moxifloxacin and proflavine in the detection of neoplastic lesions of the human gastrointestinal tract. Methods Patients with the colonic and gastric neoplastic lesions were prospectively enrolled. The lesions were biopsied with forceps or endoscopically resected. Dual fluorescence imaging was performed by using custom axially swept wide‐field fluorescence microscopy after topical moxifloxacin and proflavine instillation. Imaging results were compared with both confocal imaging with cell labeling and conventional histological examination. Results Ten colonic samples (one normal mucosa, nine adenomas) from eight patients and six gastric samples (one normal mucosa, five adenomas) from four patients were evaluated. Dual fluorescence imaging visualized detail cellular structures. Regular glandular structures with polarized cell arrangement were observed in normal mucosa. Goblet cells were preserved in normal colonic mucosa. Irregular glandular structures with scanty cytoplasm and dispersed elongated nuclei were observed in adenomas. Goblet cells were scarce or lost in the colonic lesions. Similarity analysis between moxifloxacin and proflavine imaging showed relatively high correlation values in adenoma compared with those in normal mucosa. Dual fluorescence imaging showed good detection accuracies of 82.3% and 86.0% in the colonic and the gastric lesions, respectively. Conclusions High‐contrast and high‐resolution dual fluorescence imaging was feasible for obtaining detail histopathological information in the gastrointestinal neoplastic lesions. Further studies are needed to develop dual fluorescence imaging as an in vivo real‐time visual diagnostic method.

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Two-photon microscopy of fungal keratitis-affected rabbit cornea ex vivo using moxifloxacin as a labeling agent

Cited by 9 publications

References 23 publications

Use of red, far-red, and near-infrared light in imaging of yeasts and filamentous fungi

Use of red, far-red, and near-infrared light in imaging of yeasts and filamentous fungi

A Bright Future for Fluorescence Imaging of Fungi in Living Hosts

Feasibility of moxifloxacin and proflavine dual fluorescence imaging for detecting gastrointestinal neoplastic lesions: A prospective study

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