Assessment of Extramammary Paget Disease by Two-Photon Microscopy

Abstract

Two-photon microscopy techniques are non-linear optical imaging methods which are gaining momentum in the investigation of fixed tissue sections, fresh tissue or even for in vivo experiments. Two-photon excited fluorescence and second harmonic generation are two non-linear optical contrast mechanisms which can be simultaneously used for offering complementary information on the tissue architecture. While the former can originate from endogenous autofluorescence sources (e.g., NADH, FAD, elastin, keratin, lipofuscins, or melanin), or exogenous eosin, the latter is generated in fibrillar structures within living organisms (e.g., collagen and myosin). Here we test the ability of both these contrast mechanisms to highlight features of the extramammary Paget disease on fixed tissue sections prepared for standard histological examination using immunohistochemical markers and hematoxylin and eosin staining. We also demonstrate the label-free abilities of both imaging techniques to highlight histological features on unstained fixed tissue sections. The study demonstrated that two-photon microscopy can detect specific cellular features of the extramammary Paget disease in good correlation with histopathological results.

Multiphoton microscopy assessment of the structure and variability changes of dermal connective tissue in vulvar lichen sclerosus: A pilot study

“…Such substances can be type I, III and V collagens, glycosaminoglycans, proteoglycans [25, 30, 60, 61], which actively accumulate in the dermis in VLS [62, 63]. Importantly, our results confirm Hristu et al [64], Huang et al [65] results that normal vulvar skin has TPEF signal from elastic fibers only (Figures 1C and 7H).…”

Imaging of colorectal adenomas with pseudoinvasion and malignant polyps using two-photon excitation microscopy

“…To highlight the local stromal alterations in both adenomas with epithelial misplacement and malignant polyps, we propose employing a nonlinear optical imaging technique. Specifically, we demonstrate the efficacy of two-photon excitation microscopy (TPEM) ( 16 – 19 ), a method that has gained prominence in clinical microscopy ( 20 ) in recent years and has been applied in endoscopic procedures ( 21 , 22 ). TPEM simultaneously utilizes two nonlinear optical contrast mechanisms—second harmonic generation (SHG) and two-photon excitation fluorescence (TPEF)—to acquire images.…”