Dynamic longitudinal investigation of individual nerve endings in the skin of anesthetized mice using in vivo two-photon microscopy

Journal of Biomedical Optics
Mikhail Yuryev, Leonard Khiroug

Abstract

Visualization of individual cutaneous nerve endings has previously relied on laborious procedures of tissue excision, fixation, sectioning and staining for light or electron microscopy. We present a method for non-invasive, longitudinal two-photon microscopy of single nerve endings within the skin of anesthetized transgenic mice. Besides excellent signal-to-background ratio and nanometer-scale spatial resolution, this method offers time-lapse "movies" of pathophysiological changes in nerve fine structure over minutes, hours, days or weeks. Structure of keratinocytes and dermal matrix is visualized simultaneously with nerve endings, providing clear landmarks for longitudinal analysis. We further demonstrate feasibility of dissecting individual nerve fibers with infra-red laser and monitoring their degradation and regeneration. In summary, our excision-free optical biopsy technique is ideal for longitudinal microscopic analysis of animal skin and skin innervations in vivo and can be applied widely in preclinical models of chronic pain, allergies, skin cancers and a variety of dermatological disorders.

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Citations

Apr 3, 2012·Journal of Controlled Release : Official Journal of the Controlled Release Society·Sari PappinenArto Urtti
Jul 30, 2014·Skin Research and Technology : Official Journal of International Society for Bioengineering and the Skin (ISBS) [and] International Society for Digital Imaging of Skin (ISDIS) [and] International Society for Skin Imaging (ISSI)·M J KoehlerM Kaatz
Jul 1, 2017·Frontiers in Molecular Neuroscience·Omer BarkaiAlexander M Binshtok
Apr 22, 2021·The Journal of Comparative Neurology·Han-Yuan YehChen-Tung Yen
Apr 24, 2019·The Journal of Investigative Dermatology·Malte Casper

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