In the past, significant effort has been made to develop ultrathin membranes exhibiting physiologically relevant mechanical properties, such as thickness and elasticity of native basement membranes. However, most of these fabricated membranes have a relatively high elastic modulus, ∼MPa–GPa, relevant only to retinal and epithelial basement membranes. Vascular basement membranes exhibiting relatively low elastic modulus, ∼kPa, on the contrary, have seldom been mimicked. Membranes demonstrating high compliance, with moduli ranging in ∼kPa along with sub-microscale thicknesses have rarely been reported, and would be ideal to mimic vascular basement membranes in vitro. To address this, we fabricate ultrathin membranes demonstrating the mechanistic features exhibited by their vascular biological counterparts. Salient features of the fabricated ultrathin membranes include free suspension, physiologically relevant thickness ∼sub-micrometers, relatively low modulus ∼kPa, and sufficiently large culture area ∼20 mm2. To fabricate such ultrathin membranes, undiluted PDMS Sylgard 527 was utilized as opposed to the conventional diluted polymer–solvent mixture approach. In addition, the necessity to have a sacrificial layer for releasing membranes from the underlying substrates was also eliminated in our approach. The novelty of our work lies in achieving the distinct combination of membranes having thickness in sub-micrometers and the associated elasticity in kilopascal using undiluted polymer, which past approaches with dilution have not been able to accomplish. The ultrathin membranes with average thickness of 972 nm (thick) and 570 nm (thin) were estimated to have an elastic modulus of 45 and 214 kPa, respectively. Contact angle measurements revealed the ultrathin membranes exhibited hybrophobic characteristics in unpeeled state and transformed to hydrophilic behavior when freely suspended. Human umbilical vein endothelial cells were cultured on the polymeric ultrathin membranes, and the temporal cell response to change in local compliance of the membranes was studied by evaluating the cell spread area, density, percentage area coverage, and spread rate. After 24 h, single cells, pairs, and group of three to four cells were noticed on highly compliant thick membranes, having average thickness of 972 nm and modulus of 45 kPa. On the contrary, the cell monolayer was noted on the glass slide acting as a control. For the thin membranes featuring average thickness of 570 nm and modulus of 214 kPa, the cells tend to exhibit response similar to that on control with initiation of monolayer formation. Our results indicate, the local compliance, in turn, the membrane thickness governs the cell behavior and this can have vital implications during disease initiation and progression, wound healing, and cancer cell metastasis.
Summary Commercial high‐resolution optical microscopes are essential for microscopy imaging; however, they are expensive and bulky, which limits their use in point‐of‐care devices, resource‐limited areas, and real‐time imaging of a sample in a large apparatus. In this study, we report a novel compact (10 cm × 5 cm × 5 cm, without the light source) lightweight (∼0.5 kg) submicron‐resolution inverted optical microscope at low cost (∼$ 300). Our technique utilises the proximity of the image sensor to a commercial microscope objective lens for compactness of the microscope. The use of an image sensor with a small pixel size helps to reduce the information loss, which provides high‐resolution images. Moreover, our technique offers a freedom to tailor the design of microscope according to the required resolution, cost, and portability for specific applications, which makes it a suitable candidate for affordable point‐of‐care devices. Images of several micron‐to‐submicron scale patterns and spherical beads are acquired to observe the resolution and quality of the images obtained using our microscope. In addition, we demonstrate the applications of our microscope in various fields such as recording of high‐speed water microdroplet formation inside a microfluidic device, high‐resolution live cell imaging inside an incubator, and real‐time imaging of crack propagation in a sample under stretching by a material testing system (MTS). Therefore, this portable and inexpensive microscope provides the essential functionalities of a bulky expensive high‐performance microscope at a lower cost. Lay Description Microscope is an essential tool in research allowing for observation of microsized objects and life forms. Contemporary commercial high‐resolution microscopes have long optical paths involving series of lenses and filters. Although this configuration precisely corrects for optical distortions and produces clear images, it makes modern microscopes very costly and bulky, restricting their usage to low‐funded research laboratories and at remote places. We have developed a simple digital microscope with high‐resolution but with much smaller size and lighter in weight at low cost by removing the long optical terrain. Our microscope consists of a commercial microscope objective lens for magnification and semiconductor image sensor with small pixels placed right after the lens, both of which are affordable and easily available. The small pixel size helps to translate the magnified analogue sample image to high‐resolution digital image. In our paper, we show that our microscope can view micro and submicron‐sized patterns and beads. Moreover, our fist‐sized microscope can be placed inside an incubator for real‐time imaging of cells or rotated sideways for recording submicron‐sized crack generation due stretching of novel materials, both of which could not be accomplished with the 2 feet tall laboratory microscopes.
Long-range chromosomal travel is a phenomenon unique to cell division. Methods for non-invasive, artificial manipulation of chromosomes, such as optical or magnetic tweezers, have difficulty in producing the motion of whole chromosomes in live cells. Here, we report the spatial control of chromosomes over 10 μm in a live mouse oocyte using magnetic particles driven by an external magnetic field. Selective capture of the chromosomes was achieved using antibodies specific for histone H1 in the chromosome that were conjugated to magnetic particles (H1-BMPs). When an external magnetic field was applied, the chromosomes captured by the H1-BMPs traveled through the cytosol and accumulated near the cell membrane though the movement of the chromosomes captured by H1-BMPs was strongly disturbed by the distribution of the cytoskeleton (e.g. actin filaments). Being non-invasive in nature, our approach will enable new opportunities in the remote manipulation of subcellular elements.
Stomata, functionally specialized micrometer-sized pores on the epidermis of leaves (mainly on the lower epidermis), control the flow of gases and water between the interior of the plant and atmosphere. Real-time monitoring of stomatal dynamics can be used for predicting the plant hydraulics, photosensitivity, and gas exchanges effectively. To date, several techniques offer the direct or indirect measurement of stomatal dynamics, yet none offer real-time, long-term persistent measurement of multiple stomal apertures simultaneously of an intact leaf in a field under natural conditions. Here, we report a high-resolution portable microscope-based technique for in situ real-time field imaging and monitoring of stomata. Our technique is capable of analyzing and quantifying the multiple lower epidermis stomal pore dynamics simultaneously and does not require any physical or chemical manipulation of a leaf. An upward facing objective lens in our portable microscope allows the imaging of lower epidermis stomatal opening of a leaf while upper epidermis being exposed to the natural environment. Small depth of field (~ 1.3 µm) of a high-magnifying objection lens assists in focusing the stomatal plane in highly non-planar tomato leaf having a high density of trichome (hair-like structures). For long-term monitoring, the leaf is fixed mechanically by a novel designed leaf holder providing freedom to expose the upper epidermis to the sunlight and lower epidermis to the wind simultaneously. In our study, a direct relation between the stomatal opening and the intensity of sunlight illuminating on the upper epidermis has been observed in real-time.In addition, real-time porosity of leaf (ratio between the areas of stomatal opening to the area of a leaf) and stomatal aspect ratio (ratio between the major axis and minor axis of stomatal opening) along with stomatal density have been quantified.
Metal-organic frameworks (MOFs) are arguably a class of highly tuneable polymer-based materials with wide applicability. The arrangement of chemical components and the bonds they form through specific chemical bond associations are critical determining factors in their functionality. In particular, crystalline porous materials continue to inspire their development and advancement towards sustainable and renewable materials for clean energy conversion and storage. An important area of development is the application of MOFs in proton-exchange membrane fuel cells (PEMFCs) and are attractive for efficient low-temperature energy conversion. The practical implementation of fuel cells, however, is faced by performance challenges. To address some of the technical issues, a more critical consideration of key problems is now driving a conceptualised approach to advance the application of PEMFCs. Central to this idea is the emerging field MOF-based systems, which are currently being adopted and proving to be a more efficient and durable means of creating electrodes and electrolytes for proton−exchange membrane fuel cells. This review proposes to discuss some of the key advancements in the modification of PEMs and electrodes, which primarily use functionally important MOFs. Further, we propose to correlate MOF-based PEMFC design and the deeper correlation with performance by comparing proton conductivities and catalytic activities for selected works.
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