Nonlinear Optical Microscopy: From Fundamentals to Applications in Live Bioimaging

Overview

Journal Front Bioeng Biotechnol

Date 2020 Nov 9

PMID 33163482

Citations 21

Authors

Valentina Parodi

Emanuela Jacchetti

Roberto Osellame

Giulio Cerullo

Dario Polli

Manuela Teresa Raimondi

Affiliations

Soon will be listed here.

Abstract

A recent challenge in the field of bioimaging is to image vital, thick, and complex tissues in real time and in non-invasive mode. Among the different tools available for diagnostics, nonlinear optical (NLO) multi-photon microscopy allows label-free non-destructive investigation of physio-pathological processes in live samples at sub-cellular spatial resolution, enabling to study the mechanisms underlying several cellular functions. In this review, we discuss the fundamentals of NLO microscopy and the techniques suitable for biological applications, such as two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG-THG), and coherent Raman scattering (CRS). In addition, we present a few of the most recent examples of NLO imaging employed as a label-free diagnostic instrument to functionally monitor and vital biological specimens in their unperturbed state, highlighting the technological advantages of multi-modal, multi-photon NLO microscopy and the outstanding challenges in biomedical engineering applications.

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