Abstract
Skeletal muscle is a fibrous tissue composed by muscle fibers and interstitial fluid. Due to this constitution, the muscle presents a non uniform refractive index profile that origins strong light scattering. One way to improve tissue transmittance is to reduce this refractive index mismatch by immersing the muscle in an optical clearing agent. As a consequence of such immersion tissue also suffers dehydration. The study of the optical clearing effect created by a simple mixture composed by ethanol, glycerol and distilled water has proven its effectiveness according to the variations observed in the parameters under study. The effect was characterized in terms of its magnitude, time duration and histological variations. The applied treatment has created a small reduction of the global sample refractive index that is justified by the long time rehydration caused by water in the immersing solution. From the reduction in sample pH we could also identify the dehydration process created in the sample. The immersion treatment has originated fiber bundle contraction and a spread distribution of the muscle fiber bundles inside. New studies with the mixture used, or with other combinations of its constituents might be interesting to perform with the objective to develop new clinical procedures.

Abstract
Optical characterization and internal structure of biological tissues is highly important for biomedical optics. In particular for optical clearing processes, such information is of vital importance to understand the mechanisms involved through the variation of the refractive indices of tissue components. The skeletal muscle presents a fibrous structure with an internal arrangement of muscle fiber cords surrounded by interstitial fluid that is responsible for strong light scattering. To determine the refractive index of muscle components we have used a simple method of measuring tissue mass and refractive index during dehydration. After performing measurements for natural and ten dehydration states of the muscle samples, we have determined the dependence between the refractive index of the muscle and its water content. Also, we have joined our measurements with some values reported in literature to perform some calculations that have permitted to determine the refractive index of the dried muscle fibers and their corresponding volume percentage inside the natural muscle.

Abstract
Skeletal muscle presents an internal fibrous structure. The existence of muscle fibers surrounded by interstitial fluid originates an internal step refractive index profile that causes light scattering. One way to minimize this effect inside a muscle is to perform an optical clearing treatment, using an adequate solution that presents a refractive index higher than the interstitial fluid. We have studied muscle spectral transmittance during sample immersion in propylene glycol. With the collection of transmittance spectra registered during a period of 20 minutes of immersion we could represent spectral transmittance evolution for several wavelengths and verify that the tissue samples have become more translucent. The optical clearing effect created in the tissue samples was characterized by an increase of 45% above the natural transmittance and the variations observed in tissue mass, pH and global refractive index. We also identified the initial mechanisms of agent diffusion into the tissue and consequent tissue dehydration from the spectral transmittance evolution. The histological analysis of variations caused in the internal structure of the tissues permitted to better explain the optical clearing effect created. Considering a mathematical model developed in previous studies, we could estimate the amount of agent that was inserted into the tissue samples.

Abstract
It is known that the fibrous structure of muscle causes light scattering. This phenomenon occurs due to the refractive index discontinuities located between muscle fibers and interstitial fluid. To study the possibility of reducing light scattering inside muscle, we consider its spectral transmittance evolution during an immersion treatment with an optical clearing solution containing ethanol, glycerol, and distilled water. Our methodology consists of registering spectral transmittance of muscle samples while immersed in that solution. With the spectral data collected, we represent the transmittance evolution for some wavelengths during the treatment applied. Additionally, we study the variations that the treatment has caused on the samples regarding tissue refractive index and mass. By analyzing microscopic photographs of tissue cross section, we can also verify changes in the internal arrangement of muscle fibers caused by the immersion treatment. Due to a mathematical model that we develop, we can explain the variations observed in the studied parameters and estimate the amount of optical clearing agent that has diffused into the tissue samples during the immersion treatment. At the end of the study, we observe and explain the improvement in tissue spectral transmittance, which is approximately 65% after 20 min. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3486539]

Abstract
[No abstract available]

Abstract
The optical immersion clearing is an effective method to reduce light scattering in tissues, but to optimize each treatment, it is necessary to understand the mechanisms involved. Since these treatments are intended to be temporary, it is also important to know if the mechanisms involved are reversible. Various studies have been made to evaluate and characterize the mechanisms of optical clearing. In all cases studied, two major mechanisms were observed—the tissue dehydration and the refractive index matching mechanisms. Some particular studies have reported that the agents used in treatments also dissolve proteins and suggested that protein dissolution is also a clearing mechanism. All these mechanisms have been reported as reversible, both on ex vivo or on in vivo studies. We make an analysis on these studies and present a method based on ex vivo collimated transmittance and thickness measurements to characterize the major clearing mechanisms—tissue dehydration and refractive index matching. Although this method can only be made with ex vivo tissues, alternative measurements are suggested for in vivo characterization of the clearing mechanisms. © 2019, The Author(s), under exclusive license to Springer Nature Switzerland AG.