Refractive Index Matching Medium Research Articles

Immersion laser separation: Enhancing efficiency and quality in cutting irregular lenses

Irregular lenses are commonly employed in various beam shaping, imaging, and other optical applications. They require secondary processing to meet specific design and assembly criteria. Bessel beam lasers have gained popularity for their ability to efficiently separate hard-brittle materials owing to their high processing quality and speed. However, the complex surface of irregular lenses creates challenges such as refraction and scattering, which impede the laser beam’s ability to form a laser center lobe inside the material. This paper proposes an innovative technique called immersion laser separation (ILS), where an irregular lens is immersed in a refractive index matching medium to create a compound with a uniform refractive index. This approach mitigates the impact of surface complexity on the optical path of the Bessel beam. The separation of a fast axis collimator (FAC) by ILS is used as an example to test the proposed technique. The distribution characteristics of the Bessel beam light field inside the lens material under different refractive index were investigated to find that the laser consistently forms a laser center lobe inside the material during cutting when the matching deviation is small. This results in a modified area that covers the entire surface. The maximum cutting speed can reach 50 mm/s, the cutting surface roughness is less than 700 nm, and the technique produces no chips or micro-cracks, ensuring uniformly high-quality separation. Experimental and simulation results are in close agreement, suggesting that the proposed technique is effective for the efficient, high-quality cutting of irregular lenses.

Instantaneous Clearing of Biofilm (iCBiofilm): an optical approach to revisit bacterial and fungal biofilm imaging

Whole-biofilm imaging at single-cell resolution is necessary for system-level analysis of cellular heterogeneity, identification of key matrix component functions and response to immune cells and antimicrobials. To this end, we developed a whole-biofilm clearing and imaging method, termed instantaneous clearing of biofilm (iCBiofilm). iCBiofilm is a simple, rapid, and efficient method involving the immersion of biofilm samples in a refractive index-matching medium, enabling instant whole-biofilm imaging with confocal laser scanning microscopy. We also developed non-fixing iCBiofilm, enabling live and dynamic imaging of biofilm development and actions of antimicrobials. iCBiofilm is applicable for multicolor imaging of fluorescent proteins, immunostained matrix components, and fluorescence labeled cells in biofilms with a thickness of several hundred micrometers. iCBiofilm is scalable from bacterial to fungal biofilms and can be used to observe biofilm-neutrophil interactions. iCBiofilm therefore represents an important advance for examining the dynamics and functions of biofilms and revisiting bacterial and fungal biofilm formation.

Improved 3D cellular morphometry of Caenorhabditis elegans embryos using a refractive index matching medium.

Cell shape change is one of the driving forces of animal morphogenesis, and the model organism Caenorhabditis elegans has played a significant role in analyzing the underlying mechanisms involved. The analysis of cell shape change requires quantification of cellular shape descriptors, a method known as cellular morphometry. However, standard C. elegans live imaging methods limit the capability of cellular morphometry in 3D, as spherical aberrations generated by samples and the surrounding medium misalign optical paths. Here, we report a 3D live imaging method for C. elegans embryos that minimized spherical aberrations caused by refractive index (RI) mismatch. We determined the composition of a refractive index matching medium (RIMM) for C. elegans live imaging. The 3D live imaging with the RIMM resulted in a higher signal intensity in the deeper cell layers. We also found that the obtained images improved the 3D cell segmentation quality. Furthermore, our 3D cellular morphometry and 2D cell shape simulation indicated that the germ cell precursor P4 had exceptionally high cortical tension. Our results demonstrate that the RIMM is a cost-effective solution to improve the 3D cellular morphometry of C. elegans. The application of this method should facilitate understanding of C. elegans morphogenesis from the perspective of cell shape changes.

A tunable refractive index matching medium for live imaging cells, tissues and model organisms.

In light microscopy, refractive index mismatches between media and sample cause spherical aberrations that often limit penetration depth and resolution. Optical clearing techniques can alleviate these mismatches, but they are so far limited to fixed samples. We present Iodixanol as a non-toxic medium supplement that allows refractive index matching in live specimens and thus substantially improves image quality in live-imaged primary cell cultures, planarians, zebrafish and human cerebral organoids.

Pore-Scale Flow Measurements at the Interface between a Sandy Layer and a Model Porous Medium: Application to Statistical Modeling of Contact Erosion

Contact erosion is potentially initiated at the interface between two soil layers by a groundwater flow within the coarser material. Once eroded by the flow, particles from the finer soil are transported through the pores of the coarser layer. Fluvial dikes are often exposed to this phenomenon. Small-scale experiments combining refractive index matching medium, planar laser-induced fluorescence, and particle image velocimetry were carried out to measure the flow characteristics in the vicinity of an interface between a model granular medium and a fine graded sandy layer. Longitudinal velocities and shear-stress distributions were obtained in Darcy flow conditions. They revealed a long tail toward large values, which reflects the spatial variability of the constrictions in the pores network. Taking into account these distributions can improve the modeling of contact erosion by going beyond the simple use of mean quantities, like Darcy velocity, as is usually proposed in the literature. This is done by successively considering the variability of the porous flow and also that of the critical shear stress. As a main consequence, the resulting global erosion rate is now nonzero for any value of the mean shear stress and there is no longer a stress threshold. Finally, the effect of paving at the sand surface can also be added to the statistical model and makes it possible to account very satisfactorily for previous contact erosion tests at the sample scale.

Optimization of microscope unit for studying fluorescence emitters under high-vacuum and ambient gas conditions: Optical properties for various ionic liquids as a refractive index matching medium

We briefly introduce our microscope unit with an immersion objective and ionic liquid used as a refractive index matching medium, and describe its application to single molecular spectroscopy under high-vacuum and various ambient gas conditions. In this article, we particularly focus our attention on studies toward optimizing the microscope unit. For this purpose, the refractive index and background fluorescence, absorption spectrum for various ionic liquids wre investigated. We also measured the images of the beam spot of excitation laser at focal point by using an immersion objective and various ionic liquids. We found that some have more suitable characteristics than before.

Time-Correlated Single-Photon Counting System with a Modified Objective Unit for Studying Fluorescence Emitters under High-Vacuum and Ambient Gas Conditions

In this study, we introduce a modified microscope unit for studying fluorescence emitters under high-vacuum and ambient gas conditions. The modified microscope unit has an immersion objective and uses an ionic liquid as a refractive index matching medium. In our optical geometry, the immersion objective and ionic liquid are placed in high vacuum and ambient gas. The optical properties of single colloidal quantum dots (CdSe/ZnS) in a poly(methyl methacrylate) (PMMA) matrix placed in ambient air, high vacuum, and ambient nitrogen gas with an appropriate pressure are studied.

Time-correlated single photon counting system and light-collection system for studying fluorescence emitters under high-vacuum conditions: Use of immersion objective and ionic liquid

In this paper, we introduce a time-correlated single photon counting system and an efficient light-collection system for studying fluorescence emitters under high-vacuum conditions; the latter system has an immersion objective, and an ionic liquid is used as a refractive index matching medium. The ionic liquid is positioned in high vacuum. The time-correlated single photon counting system has modified photomultiplier tubes that act as photon detectors. The light-collection system is designed to be simple, compact, and easy to use. In order to verify the performance of these systems, the optical properties of colloidal semiconductor nanocrystals (CdSe/ZnS) and fluorescent dye molecules are studied as examples.