In vivo characterization of minipig skin as a model for dermatological research using multiphoton microscopy

Alex, Aneesh; Chaney, Eric J.; Žurauskas, Mantas; Criley, Jennifer M; Spillman, Darold R.; Hutchison, Phaedra B.; Li, Joanne; Marjanović, Marina; Frey, Steve; Arp, Zane; Boppart, Stephen A.

doi:10.1111/exd.14152

Cited by 20 publications

(12 citation statements)

References 41 publications

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“…We had previously demonstrated in vivo FLIM imaging of murine skin using a typical TCSPC-based setup which took 51 s per frame, compared to the 2.5 s (and real-time processing) for FOCALS microscopy 54 . FOCALS microscopy also compares favorably against commercial FLIM imaging systems previously used for in vivo skin imaging 55 , 56 with a 5–6 × improvement in imaging speed, respectively.…”

Section: Discussionmentioning

confidence: 99%

Label-free metabolic and structural profiling of dynamic biological samples using multimodal optical microscopy with sensorless adaptive optics

Iyer

Sorrells

Yang

et al. 2022

Sci Rep

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Label-free optical microscopy has matured as a noninvasive tool for biological imaging; yet, it is criticized for its lack of specificity, slow acquisition and processing times, and weak and noisy optical signals that lead to inaccuracies in quantification. We introduce FOCALS (Fast Optical Coherence, Autofluorescence Lifetime imaging, and Second harmonic generation) microscopy capable of generating NAD(P)H fluorescence lifetime, second harmonic generation (SHG), and polarization-sensitive optical coherence microscopy (OCM) images simultaneously. Multimodal imaging generates quantitative metabolic and morphological profiles of biological samples in vitro, ex vivo, and in vivo. Fast analog detection of fluorescence lifetime and real-time processing on a graphical processing unit enables longitudinal imaging of biological dynamics. We detail the effect of optical aberrations on the accuracy of FLIM beyond the context of undistorting image features. To compensate for the sample-induced aberrations, we implemented a closed-loop single-shot sensorless adaptive optics solution, which uses computational adaptive optics of OCM for wavefront estimation within 2 s and improves the quality of quantitative fluorescence imaging in thick tissues. Multimodal imaging with complementary contrasts improves the specificity and enables multidimensional quantification of the optical signatures in vitro, ex vivo, and in vivo, fast acquisition and real-time processing improve imaging speed by 4–40 × while maintaining enough signal for quantitative nonlinear microscopy, and adaptive optics improves the overall versatility, which enable FOCALS microscopy to overcome the limits of traditional label-free imaging techniques.

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Section: Discussionmentioning

confidence: 99%

Label-free metabolic and structural profiling of dynamic biological samples using multimodal optical microscopy with sensorless adaptive optics

Iyer

Sorrells

Yang

et al. 2022

Sci Rep

Self Cite

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“…It thus provides more information that may help to understand topically applied exogenous molecules diffusion, intradermal distribution, and penetration depth (25,26). FLIM tomography can be used for the investigation of their impact on the metabolism of skin cells directly in vivo (27)(28)(29).…”

Section: Discussionmentioning

confidence: 99%

In vivo two-photon imaging of topically applied care cosmetics and pharmaceuticals in human skin

König

2023

Multiphoton Microscopy in the Biomedical Sciences XXIII

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We applied femtosecond laser multiphoton tomographs, including the novel system MPTcompact with its chiller-free fiber laser positioned inside the measurement head, for multimodal imaging of in vivo human skin of volunteers, that had been exposed to topically applied sunscreens, UV stains, care cosmetics, and pharmaceuticals. Signals are based on two-photon excited and time-resolved fluorescence (FLIM), Second Harmonic Generation (SHG), and confocal reflection. In particular, SHG signals have been detected from silver nanoparticles in mineral sunscreen. Of particular interest was the distribution of the exogenous compounds within the upper epidermis such as the stratum corneum as an efficient skin barrier. The lateral resolution was 300 nm and the time resolution was 250 picoseconds. In addition, the autofluorescence based on NADH and keratin detection was imaged.

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“…No information has been found in minipigs, despite their increasing use in different branches of translational medicine [140][141][142][143]. Further specific studies should be performed characterizing the effects induced by anaesthetic drugs in these animals, also considering that most of them are used as adult.…”

Section: Plos Onementioning

confidence: 99%

How is depth of anaesthesia assessed in experimental pigs? A scoping review

et al. 2023

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Background Despite the large number of pigs involved in translational studies, no gold standard depth of anaesthesia indicators are available. We undertook a scoping review to investigate and summarize the evidence that sustains or contradicts the use of depth of anaesthesia indicators in this species. Methods Medline, Embase and CAB abstract were searched up to September 22nd 2022. No limits were set for time, language and study type. Only original articles of in vivo studies using pigs or minipigs undergoing general anaesthesia were included. The depth of anaesthesia indicators reported in the selected papers were divided in two categories: A, indicators purposely investigated as method to assess depth of anaesthesia; B, indicators reported but not investigated as method to assess depth of anaesthesia. Results Out of 13792 papers found, 105 were included after the screening process. Category A: 17 depth of anaesthesia indicators were found in 19 papers. Studies were conducted using inhalant anaesthetics as the main anaesthetic agent in the majority of the cases (13/19 = 68.4%), while 3/19 (15.8%) used propofol. The most investigated depth of anaesthesia indicators were bispectral index (8/19 = 42.1%) and spectral edge frequency 95% (5/19 = 26.3%). Contrasting results about the specific usefulness of each depth of anaesthesia indicators were reported. Category B: 23 depth of anaesthesia indicators were found in 92 papers. The most reported depth of anaesthesia indicators were: motor response following a stimulus (37/92 = 40.2%), depth of anaesthesia scores (21/92 = 23.3%), bispectral index (16/92 = 17.8%) and spectral edge frequency 95% (9/92 = 9.8%). Conclusion Results highlight the lack of scientifically valid and reliable indicators to ensure adequate depth of anaesthesia in pigs.

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In vivo characterization of minipig skin as a model for dermatological research using multiphoton microscopy

Cited by 20 publications

References 41 publications

Label-free metabolic and structural profiling of dynamic biological samples using multimodal optical microscopy with sensorless adaptive optics

Label-free metabolic and structural profiling of dynamic biological samples using multimodal optical microscopy with sensorless adaptive optics

In vivo two-photon imaging of topically applied care cosmetics and pharmaceuticals in human skin

How is depth of anaesthesia assessed in experimental pigs? A scoping review

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