No abstract

Sample preparation is critical to biological electron microscopy (EM), and there have been continuous efforts on optimizing the procedures to best preserve structures of interest in the sample. However, a quantitative characterization of the morphological changes associated with each step in EM sample preparation is currently lacking. Using correlative EM and superresolution microscopy (SRM), we have examined the effects of different drying methods as well as osmium tetroxide (OsO4) post-fixation on cell morphology during scanning electron microscopy (SEM) sample preparation. Here, SRM images of the sample acquired under hydrated conditions were used as a baseline for evaluating morphological changes as the sample went through SEM sample processing. We found that both chemical drying and critical point drying lead to a mild cellular boundary retraction of ~60 nm. Post-fixation by OsO4 causes at least 40 nm additional boundary retraction. We also found that coating coverslips with adhesion molecules such as fibronectin prior to cell plating helps reduce cell distortion from OsO4 post-fixation. These quantitative measurements offer useful information for identifying causes of cell distortions in SEM sample preparation and improving current procedures.

No abstract

Purpose There are over 40 million amputees globally with more than 185,000 Americans losing their limbs every year. For most of the world, prosthetic devices remain too expensive and uncomfortable. This paper aims to outline advancements made by a multidisciplinary research group, interested in advancing the restoration of human motion through accessible lower limb prostheses. Design/methodology/approach Customization, comfort and functionality are the most important metrics reported by prosthetists and patients. The work of this paper presents the design and manufacturing of a custom made, cost-effective and functional three-dimensional (3D) printed transtibial prosthesis monocoque design. The design of the prosthesis integrates 3D imaging, modelling and optimization techniques coupled with additive manufacturing. Findings The successful fabrication of a functional monocoque prosthesis through 3D printing indicates the workflow may be a solution to the worldwide accessibility crisis. The digital workflow developed in this work offers great potential for providing prosthetic devices to rural communities, which lack access to skilled prosthetic physicians. The authors found that using the workflow together with 3D printing, this study can create custom monocoque prostheses (Figure 16). These prostheses are comfortable, functional and properly aligned. In comparison with traditional prosthetic devices, the authors slowered the average cost, weight and time of production by 95%, 55% and 95%, respectively. Social implications This novel digital design and manufacturing workflow has the potential to democratize and globally proliferate access to prosthetic devices, which restore the patient’s mobility, quality of life and health. LIMBER’s toolbox can reach places where proper prosthetic and orthotic care is not available. The digital workflow reduces the cost of making custom devices by an order of magnitude, enabling broader reach, faster access and improved comfort. This is particularly important for children who grow quickly and need new devices every few months or years, timely access is both physically and psychologically important. Originality/value In this manuscript, the authors show the application of digital design techniques for fabricating prosthetic devices. The proposed workflow implements several advantageous changes and, most importantly, digitally blends the three components of a transtibial prosthesis into a single, 3D printable monocoque device. The development of a novel unibody transtibial device that is properly aligned and adjusted digitally, greatly reduces the number of visits an amputee must make to a clinic to have a certified prosthetist adjust and modify their prosthesis. The authors believe this novel workflow has the potential to ease the worldwide accessibility crisis for prostheses.