Multimodal non-linear microscopy combining coherent anti-Stokes Raman scattering, second harmonic generation, and two-photon excited fluorescence has proved to be a versatile and powerful tool enabling the label-free investigation of tissue structure, molecular composition, and correlation with function and disease status. For a routine medical application, the implementation of this approach into an in vivo imaging endoscope is required. However, this is a difficult task due to the requirements of a multicolour ultrashort laser delivery from a compact and robust laser source through a fiber with low losses and temporal synchronization, the efficient signal collection in epi-direction, the need for small-diameter but highly corrected endomicroobjectives of high numerical aperture and compact scanners. Here, we introduce an ultra-compact fiber-scanning endoscope platform for multimodal non-linear endomicroscopy in combination with a compact four-wave mixing based fiber laser. The heart of this fiber-scanning endoscope is an in-house custom-designed, single mode, double clad, double core pure silica fiber in combination with a 2.4 mm diameter NIR-dual-waveband corrected endomicroscopic objective of 0.55 numerical aperture and 180 µm field of view for non-linear imaging, allowing a background free, low-loss, high peak power laser delivery, and an efficient signal collection in backward direction. A linear diffractive optical grating overlays pump and Stokes laser foci across the full field of view, such that diffraction-limited performance is demonstrated for tissue imaging at one frame per second with sub-micron spatial resolution and at a high transmission of 65% from the laser to the specimen using a distal resonant fiber scanner.

Measuring the shape (coordinates x, y, z ) and spectral characteristics (wavelength-dependent reflectance R (λi)) of macroscopic objects as a function of time (t) is of great interest in areas such as medical imaging, precision agriculture, or optical sorting. Here, we present an approach that allows to determine all these quantities with high resolution and accuracy, enabling measurement in five dimensions. We call this approach 5D hyperspectral imaging. We describe the design and implementation of a 5D sensor operating in the visible to near-infrared spectral range, which provides excellent spatial and spectral resolution, great depth accuracy, and high frame rates. The results of various experiments strongly indicate the great benefit of the new technology.

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Here, we present a new handheld multiphoton endomicroscopic system designed for tumor diagnosis in the head and neck region. It consists of an approximate 25-cm-long rigid endomicroscopic probe with two variants (0° and 45° bended tip), connected to a handheld scan-head. The system can achieve a field of view ⪆600 µm for Coherent Anti-stokes Raman Scattering (CARS) and other nonlinear imaging modalities by a non-de-scanned detection and using a de-scanned confocal imaging channel to detect light from tissue labeled with Indocyanine Green (ICG). Furthermore, high-power femtosecond laser pulses can be transmitted through the system for precise tissue ablation which was considered in the optical design of the probe.