Optical scanning microscopy techniques based on the polarization control of the light have the capability of providing non invasive label-free contrast. By comparing the polarization states of the excitation light with its transformation after interaction with the sample, the full optical properties can be summarized in a single 4×4 Mueller matrix. The main challenge of such a technique is to encode and decode the polarized light in an optimal way pixel-by-pixel and take into account the polarimetric artifacts from the optical devices composing the instrument in a rigorous calibration step. In this review, we describe the different approaches for implementing such a technique into an optical scanning microscope, that requires a high speed rate polarization control. Thus, we explore the recent advances in term of technology from the industrial to the medical applications.

We present a new type of dual optical frequency comb source capable of scaling applications to high measurement speeds while combining high average power, ultra-low noise operation, and a compact setup. Our approach is based on a diode-pumped solid-state laser cavity which includes an intracavity biprism operated at Brewster angle to generate two spatially-separated modes with highly correlated properties. The 15-cm-long cavity uses an Yb:CALGO crystal and a semiconductor saturable absorber mirror as an end mirror to generate more than 3 W average power per comb, below 80 fs pulse duration, a repetition rate of 1.03 GHz, and a continuously tunable repetition rate difference up to 27 kHz. We carefully investigate the coherence properties of the dual-comb by a series of heterodyne measurements, revealing several important features: (1) ultra-low jitter on the uncorrelated part of the timing noise; (2) the radio frequency comb lines of the interferograms are fully resolved in free-running operation; (3) we validate that through a simple measurement of the interferograms we can determine the fluctuations of the phase of all the radio frequency comb lines; (4) this phase information is used in a post-processing routine to perform coherently averaged dual-comb spectroscopy of acetylene (C2H2) over long timescales. Our results represent a powerful and general approach to dual-comb applications by combining low noise and high power operation directly from a highly compact laser oscillator.

Circular intensity differential scattering (CIDS) is based on the analysis of circular polarized light scattering and has been proven to be an interesting label-free microscopy technique sensitive to the chiral organization at the submicroscopic level. However, this approach averages the localized contrasts related to the sample polarimetric properties in the illumination volume. Additionally, the detection sensitivity suffers from the confinement of the mixture of structures, and it becomes an arduous task to discriminate the source of the signal. In this work, we show that a phasor map approach combined with CIDS microscopy has provided an intuitive view of the sample organization to recognize the presence of different molecular species in the illumination volume. The data represented in terms of polarization response mapped to a single point called a phasor also have the potential to pave the way for the analysis of large data sets. We validated this method by numerical simulations and compared the results with that of experimental data of optical devices of reference.

Mueller Matrix Microscopy exploits the generation and the analysis of polarized light to create label-free contrast in biological images. However, when dealing with Optical Scanning Microscopy, it is required a fast generation of the polarization states in order to obtain a good Signal-to-Noise Ratio at the pixel-dwell time rate. In this work, we propose a microscopy system based on a scanning beam architecture that is exploiting the simultaneous emission of orthogonal polarization states from a Zeeman laser to provide Mueller Matrix images. This approach is based on the detection of an interference signal that allows to time-encode polarization states directly from the laser source, without the need for further active components for the management of the polarization states. We provide the theoretical model behind this approach and we apply our new method to the imaging of biological samples. Our Mueller Matrix imaging setup enables high-speed scanning microscopy, while preserving compactness and simplicity of construction. Our findings may lead to more effective dissemination of label-free techniques and their use by biological researchers.

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We demonstrate a single-cavity dual-comb laser combining 1-GHz repetition rate, 3-W average power, and 78-fs pulses via an intracavity biprism. The laser’s low noise properties enable a fully coherent dual-comb spectroscopy measurement on acetylene.

In this work, we exploited the dual-frequency, dual-polarization emission of a Zeeman laser acting as illumination stage of a multimodal optical scanning microscope to obtain polarization-resolved images of biological samples.

Polarized light scanning microscopy is a non-invasive and contrast-enhancing technique to investigate anisotropic specimens and chiral organizations. However, such arrangements suffer from insensitivity to confined blend of structures at sub-diffraction level. Here for the first time, we present that the pixel-by-pixel polarization modulation converted to an image phasor approach issues an insightful view of cells to distinguish anomalous subcellular organizations. To this target, we propose an innovative robust way for identifying changes in the chromatin compaction and distortion of nucleus morphology induced by the activation of the lamin-A gene from Hutchinson–Gilford progeria syndrome that induces a strong polarization response. The phasor mapping is evaluated based on the modulation and phase image acquired from a scanning microscope compared to a confocal fluorescence modality of normal cell opposed to the progeria. The method is validated by characterizing polarization response of starch crystalline granules. Additionally, we show that the conversion of the polarization-resolved images into the phasor could further utilized for segmenting specific structures presenting various optical properties under the polarized light. In summary, image phasor analysis offers a distinctly sensitive fast and easy representation of the polarimetric contrast that can pave the way for remote diagnosis of pathological tissues in real-time.