SHG imaging and quantitative analysis of bone collagen fibers using a multiphoton microscope
Bone is a hard tissue that supports the body, and the ratio of inorganic components (calcium etc.) in bone is only 45% by weight, with the remainder being composed of water and organic components consisting primarily of collagen. In mineralized bone, inorganic crystals mainly composed of calcium and phosphoric acid are deposited on collagen fibers, which are bundled collagen molecules. Robust and tough bone is hard and flexible. The hardness is due to inorganic components, and the flexibility is due to collagen. Prof. Tadahiro IIMURA and Dr. Takanori SATO of the Department of Pharmacology, Faculty and Graduate School of Dental Medicine, Hokkaido University are studying the pharmacological effects of PTH (teriparatide) preparation, a drug developed for the treatment of osteoporosis. This application note introduces an example of quantitative evaluation of the arrangement of collagen fibers in bone, by combining SHG (Second Harmonic Generation) imaging using a multiphoton microscope and AI technology.
The Effects of Refractive Index Mismatch Between Immersion Liquid and Tissue Clearing Reagent on Image Resolution in Deep Areas
Purkinje cells, which have a characteristic shape, are difficult to capture with a confocal system using visible light excitation, from the surface to the deep regions of the cerebellum, even if they are subjected to tissue clearing treatment, so observation with a multiphoton confocal system by IR light excitation is more suitable. Nevertheless, especially during detailed observations in deep areas, the difference between the refractive index of the immersion liquid of the objective lens and that of the tissue affects brightness and resolution. This article focuses on the effect this difference in refractive index has at different observation depths. In addition, we captured the state of the PSF along the optical axis for each depth using fluorescent beads, conducted nonlinear deconvolution processing using this result as a parameter, and verified its beneficial effect on image elongation along the optical axis.
Development of Technology to Visualize Interaction of Osteoblasts and Osteoclasts in Living Bone Tissue
A research group led by Professor Masaru Ishii (Immune cell biology), Graduate School of Medicine, Osaka University, has developed a technology to observe the inside of living bones, and to simultaneously visualize osteoblasts, which make new bone, and osteoclasts, which dissolve old bone, using the A1R MP+ multi-photon confocal microscope, which is capable of deep tissue imaging. This application note introduces an example of the world’s first successful imaging of the moment of direct contact and communication between osteoblasts and osteoclasts in living bone tissue.
The Secret Inside Flowers - Imaging Inside Plants Using ClearSee Clearing Reagent
In this Application Note, we introduce examples of clearing the flowers of Arabidopsis thaliana with ClearSee and observing them with a multiphoton excitation confocal microscope suited for observation of deep parts of organisms. This enables us to observe in detail how each of the pollen tubes elongate in the pistil for fertilization, using the color-coding of the pollen tubes.
Nikon’s Large-Format Multiphoton System for Intravital Imaging
Nikon’s A1R MP multiphoton confocal microscope system provides a choice platform for deep tissue imaging. Researchers at the Allen Institute for Brain Science have applied the A1R MP towards fast dual-color intravital imaging of the mouse brain. The open architecture instrument design allows for the addition of custom stages for large samples, external photostimulation devices, and more – perfect for constantly evolving needs.