Circular dichroism imaging has proved a powerful and simple method for extracting information on chiral molecules without specific fluorescent labels. Numerous mathematical models show that outside the absorption band, the circular dichroism signal comes from the scattering interaction and brings additional information about the organization of biopolymers. With this article, we propose a fast method to control the polarization states without moving parts, by means of a photoelastic modulator. We implemented the technique on a modified commercial confocal microscope realizing a multimodal configuration. We demonstrate its imaging capabilities by studying the organization of chromatin DNA inside isolated cell nuclei.

Zebrafish are powerful animal models for understanding biological processes and the molecular mechanisms involved in different human diseases. Advanced optical techniques based on fluorescence microscopy have become the main imaging method to characterize the development of these organisms at the microscopic level. However, the need for fluorescence probes and the consequent high light doses required to excite fluorophores can affect the biological process under observation including modification of metabolic function or phototoxicity. Here, without using any labels, we propose an implementation of a Mueller-matrix polarimeter into a commercial optical scanning microscope to characterize the polarimetric transformation of zebrafish preserved at different embryonic developmental stages. By combining the full polarimetric measurements with statistical analysis of the Lu and Chipman mathematical decomposition, we demonstrate that it is possible to quantify the structural changes of the biological organization of fixed zebrafish embryos and larvae at the cellular scale. This convenient implementation, with low light intensity requirement and cheap price, coupled with the quantitative nature of Mueller-matrix formalism, can pave the way for a better understanding of developmental biology, in which label-free techniques become a standard tool to study organisms.

A full Mueller polarimeter was implemented on a commercial laser-scanning microscope. The new polarimetric microscope is based on high-speed polarization modulation by spectral coding using a wavelength-swept laser as a source. Calibration as well as estimation of the measurement errors of the device are reported. The acquisition of Mueller images at the speed of a scanning microscope is demonstrated for the first time. Mueller images of mineral and biological samples illustrate this new polarimetric microscopy.

Expansion microscopy (ExM) is a novel preparation method enhancing the optical resolution by expanding uniformly the relative distance between fluorescence molecules on a sample placed inside a polymerized gel matrix. However, a skilled operator is needed for fluorescent labeling protocols and a high light dose is required for measurement. In this work, we couple ExM with a label-free differential circular polarization microscopy technique, demonstrated to be sensitive to the chiral organization of biopolymers. We show that by improving the distance between chiral groups, the new imaging contrast gives access to a better resolution of the chromatin-DNA organization in situ.

Calculation of the eigenvectors of two- and three-dimensional coherency matrices, and the four-dimensional coherency matrix associated with a Mueller matrix, is considered, especially for algebraic cases, in the light of recently published algorithms. The preferred approach is based on a combination of an evaluation of the characteristic polynomial and an adjugate matrix. The diagonal terms of the coherency matrix are given in terms of the characteristic polynomial of reduced matrices as functions of the eigenvalues of the coherency matrix. The analogous polynomial form for the off-diagonal elements of the coherency matrix is also presented. Simple expressions are given for the pure component in the characteristic decomposition.

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.

Measures of purity for 3D partially polarized fields, and in particular, the separation into circularly and linearly polarized contributions, are reexamined, and a new degree of total linear polarization introduced. Explicit expressions for the characteristic decomposition in terms of coherency matrix elements are presented, including the special case of an intrinsic coherency matrix. Parameterization of the coherency matrix in terms of ellipticity, and the directions of the ellipse normal and major axis are investigated. Phase consistency is discussed. A comprehensive collection of results regarding intrinsic polarization properties is presented.

A new setup is proposed to perform high-speed Mueller polarimetry by spectral coding of polarization in a reflection configuration. The system uses a swept laser source and a photodiode, which results in a simple optical setup that allows measurement of Mueller matrices at 100 kHz repetition rate. A special focus is made on the influence of the cube beam splitter polarimetric response, which is essential to measurements in a reflection configuration. The instrument is first validated on reference samples for single-point measurements, and the effect of a proper system calibration is also demonstrated on polarimetric images. The device is intended to be implemented within a laser scanning microscope to perform multimodal imaging (confocal/multiphoton and Mueller polarimetry).