Zebrafish structural development in Mueller-matrix scanning microscopy

Abstract

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.

Review on Complete Mueller Matrix Optical Scanning Microscopy Imaging

“…The main application was the study of starch molecules under certain conditions (thermal and mechanical stress). Similarly, a similar approach has been extended for small animal embryonic development analysis in the reflection configuration [45].…”

“…Furthermore, Figure 6ii shows that this discrimination can be done on the image extracted from the retardance orientation (i.e., birefringent fast axis) where it is possible to follow the rotation of the fiber in space with high imaging contrast. It could be of interest in the case the experiment requires the tracking of a specific polarimetric change under certain conditions as it has be done for fixed zebrafish reported in Figure 6iii [45]. This work has proposed to acquire a detailed fingerprint at different developmental stages and interpret all the MM parameters.…”

Polarimetric optical scanning microscopy of zebrafish embryonic development using the coherency matrix

“…In our previous work, we focused on the gold standard decomposition proposed by Lu and Chipman, such as diattenuation and retardance to highlight the polarimetric changes during the embryonic development in zebrafish, between 4 (hours post fertilization) and 72 hpf [31]. Here, from the images acquired with our experimental setup in a reflection scanning microscopy configuration, we propose a study of the specific influence of the structural biological changes on only scattering and depolarization.…”

Circular Intensity Differential Scattering for Label-Free Chromatin Characterization: A Review for Optical Microscopy

“…Advances in the remote sensing research field have allowed Mueller polarimeters to focus on the target detection of micro-/nano-particles in a turbid or highly scattering medium [ 11 , 12 , 13 , 14 , 15 ]. The main recent advances have been focused on investigating the modification of the tissues induced by pathologies at the cellular level [ 16 , 17 , 18 , 19 , 20 , 21 , 22 ] for biomedical diagnosis and on developing innovative optical label-free polarimetric microscopes [ 23 , 24 , 25 ].…”