Modified inverted selective plane illumination microscopy for sub-micrometer imaging resolution in polydimethylsiloxane soft lithography devices

Moldable, transparent polydimethylsiloxane (PDMS) elastomer microdevices enable a broad range of complex studies of three-dimensional cellular networks in their microenvironment in vitro. However, the uneven distribution of refractive index change, external to PDMS devices and internally in the sample chamber, creates a significant optical path difference (OPD) that distorts the light sheet beam and so restricts diffraction limited performance. We experimentally showed that an OPD of 120 μm results in the broadening of the lateral point spread function by over 4-fold. In this paper, we demonstrate steps to adapt a commercial inverted selective plane illumination microscope (iSPIM) and remove the OPD so as to achieve sub-micrometer imaging ranging from 0.6 ± 0.04 μm to 0.91 ± 0.03 μm of a fluorescence biological sample suspended in regular saline (RI ≈1.34) enclosed in 1.2 to 2 mm thick micromolded PDMS microdevices. We have proven that the removal of the OPD from the external PDMS layer by refractive index (RI) matching with a readily accessible, inexpensive sucrose solution is critical to achieve a >3-fold imaging resolution improvement. To monitor the RI matching process, a single-mode fiber (SMF) illuminator was integrated into the iSPIM. To remove the OPD inside the PDMS channel, we used an electrically tunable lens (ETL) that par-focuses the light sheet beam with the detection objective lens and so minimised axial distortions to attain sub-micrometer imaging resolution. We termed this new light sheet imaging protocol as modified inverted selective plane illumination microscopy (m-iSPIM). Using the high spatial–temporal 3D imaging of m-iSPIM, we experimentally captured single platelet (≈2 μm) recruitment to a platelet aggregate (22.5 μm × 22.5 μm × 6 μm) under flow at a 150 μm depth within a microfluidic channel. m-iSPIM paves the way for the application of light sheet imaging to a wide range of 3D biological models in microfluidic devices which recapitulate features of the physiological microenvironment and elucidate subcellular responses.This work was supported by grants Australian Research Council (DP200100364, DP190100039, DE160100843) and ANU Major Equipment grant (15MEC36, 16MEC26) and additional funds from ARC Centre of Excellence Translational Photosynthesis

Holo-UNet: hologram-to-hologram neural network restoration for high fidelity low light quantitative phase imaging of live cells

Intensity shot noise in digital holograms distorts the quality of the phase images after phase retrieval, limiting the usefulness of quantitative phase microscopy (QPM) systems in long term live cell imaging. In this paper, we devise a hologram-to-hologram neural network, Holo-UNet, that restores high quality digital holograms under high shot noise conditions (sub-mW/cm2 intensities) at high acquisition rates (sub-milliseconds). In comparison to current phase recovery methods, Holo-UNet denoises the recorded hologram, and so prevents shot noise from propagating through the phase retrieval step that in turn adversely affects phase and intensity images. Holo-UNet was tested on 2 independent QPM systems without any adjustment to the hardware setting. In both cases, Holo-UNet outperformed existing phase recovery and block-matching techniques by ∼ 1.8 folds in phase fidelity as measured by SSIM. Holo-UNet is immediately applicable to a wide range of other high-speed interferometric phase imaging techniques. The network paves the way towards the expansion of high-speed low light QPM biological imaging with minimal dependence on hardware constraints.Australian Research Council (DE160100843, DP190100039, DP200100364

Exploring the Collaborative Enabling and Value Innovation Mechanism in Platform Enterprises:A Case Study

The digital innovative evolution of platform enterprises has shown obvious advantanges in promoting the development of value creation, and encouraging business transformation in current digital economy era. However, we still know little about the value innovation mechanism in platform enterprises. Thus, based on grounded theory, this paper explores the characteristics of collaborative enabling and extracts three attributes -openness, symbiosis and collaboration- in driving the internal development of platform enterprises, by coding the relevant information of Shi Wai Tao Yuan (SWTY). Through the case analysis on SWTY, this paper proposes three enabling functions in the field of culture, technology and data, and builds the multi-level path of value innovation including value co-creation, resource sharing and idea symbiosis. Finally, this paper constructs a theoretical framework on collaborative enabling and value innovation in platform enterprises based on nine core categories. Overall, this paper contributes to research on the formation process and action mechanism of collaborative enabling and value innovation in platform enterprises, also shed light on the exploration on platform enterprises, and business ecosystems evolution. In doing so, this paper also provides novel insights into the sustainable development of platform enterprises in practice

Four Cases of the Rare Skin Manifestation: Monoclonal-Proteinemia

The skin manifestations of monoclonal(M)-proteinemia are rare and present in patients with monoclonal gammopathy of undetermined significance (MGUS), smoldering plasma cell myeloma (SMM) and multiple myeloma (MM). In this study, we reported 4 cases with M-proteinemia-related rare skin lesions, including pyoderma gangrenosum (PG), erythema elevatum diutinum (EED), cutis laxa (CL) and lichen myxedematosus(LM). These skin lesions are specific, where the potential mechanism was immune-mediated paraneoplastic syndrome rather than direct plasma cell infiltration. Anti-plasma cell treatment was effective in treating skin lesions. The clinical outcome of MM-related skin changes was correlated to tumor control, whereas the prognosis of MGUS or SMM related skin lesions was favorable. Skin involvement in M-proteinemia is extremely rare and less well-known, which greatly impairs quality of life. The diagnosis and treatment of these 4 cases support the need for futher study

Design of helical groove/hollow nanofibers via tri-fluid electrospinning

Electrospinning of multilevel structured micro/nanofibers have obtained considerable attention recently. In this study, inspired by the unique properties and functions of helical, groove and hollow structures in nature, we prepared a new type of multilevel structured nanofibers with helical and groove/hollow structures. Based on the fiber formation mechanism, a combination of cellulose acetate and other appropriate polymers were applied to generate helical groove/hollow nanofibers using a designed tri-fluid electrospinning system. By varying the capillary dispositions of the spinneret, the fibers with helical groove and helical hollow structures were obtained. To explore the forming process of the composite fibers, high speed photography was employed to record the Taylor cone and jet path under the spinnerets. By characterizing the morphologies and specific functions of the as-prepared nanofibers, the unique helical groove/hollow structure were generated and present good properties in wettability and mechanical behavior

STUDY OF CALCIPOTRIOL BETAMETHASONE OINTMENT IN THE TREATMENT OF PATIENTS WITH REFRACTORY CHRONIC ECZEMA

Chronic eczema is an inflammatory-immune disease of the skin, with the characteristics of skin thickening and varying degrees of lichenification, including severe itching, tendency of persistence and recurrence with serious impact on quality of life of patients. Objective: To assess the clinical efficacy and safety of calcipotriol betamethasone ointment in patients with chronic eczema. Methods: In this multi-center, randomized, single-blind, positive drug parallel controlled clinical study, patients were randomly divided into treatment and control groups, to receive calcipotriol betamethasone ointment or halometasone/triclosan cream, respectively, once daily in the evening over a 4-week period. The safety and efficacy of the two regimens were followed up on weeks 1, 2, 4 and at a 4-week treatment-free period. According to the degree of improvement, the total scores (0-4) before and after treatment and the efficacy index were calculated. The overall efficacy was assessed by four levels of evaluation model. Results: After 4 weeks of treatment, the cure rate was high (44.70%) in treatment group compared with control group (15.56%) (P<0.001), and the effective rate was 83.33% and 55.56% in the respective groups (P<0.001). At 2 and 4 weeks after treatment, there was significant difference (P<0.05) between two groups, with a reduction in the intensity of pruritus, inflammation, infiltration/ hypertrophy, lichenification, and area of target lesions. The incidence of adverse events was more (1.52%) in treatment than control group (0.00%) (P>0.05). Conclusion: Calcipotriol betamethasone ointment appears to be a safe and effective option for the treatment of chronic eczema

Coherent Optical Scattering and Interferometry (COSI) Microscopy for Morphological Imaging of Thrombus

In this work, we propose a label-free COSI system to quantify morphological changes and platelet activity along non-patterned collagen fibers within millisecond in microfluidics channels under flow at sub-platelet imaging resolution

Label-free multimodal quantitative imaging flow assay for intrathrombus formation in vitro

Microfluidics in vitro assays recapitulate a blood vessel microenvironment using surface-immobilized agonists under biofluidic flows. However, these assays do not quantify intrathrombus mass and activities of adhesive platelets at the agonist margin and use fluorescence labeling, therefore limiting clinical translation potential. Here, we describe a label-free multimodal quantitative imaging flow assay that combines rotating optical coherent scattering microscopy and quantitative phase microscopy. The combined imaging platform enables real-time evaluation of membrane fluctuations of adhesive-only platelets and total intrathrombus mass under physiological flow rates in vitro. We call this multimodal quantitative imaging flow assay coherent optical scattering and phase interferometry (COSI). COSI records intrathrombus mass to picogram accuracy and shape changes to a platelet membrane with high spatial-temporal resolution (0.4 μm/s) under physiological and pathophysiological fluid shear stress (1800 and 7500 s−1). With COSI, we generate an axial slice of 4 μm from the coverslip surface, approximately equivalent to the thickness of a single platelet, which permits nanoscale quantification of membrane fluctuation (activity) of adhesive platelets during initial adhesion, spreading, and recruitment into a developing thrombus (mass). Under fluid shear, pretreatment with a broad range metalloproteinase inhibitor (250 μM GM6001) blocked shedding of platelet adhesion receptors that shown elevated adhesive platelet activity at average of 42.1 μm/s and minimal change in intrathrombus mass.This work was supported by grants from the Australian Research Council (DP200100364, DP190100039, and DE160100843) and ANU Major Equipment grant (15MEC36 and 16MEC26)

Red Blood Cell Distribution Width: A Risk Factor for Prognosis in Patients with Ischemic Cardiomyopathy after Percutaneous Coronary Intervention

Background: It has been demonstrated in previous studies that red blood cell distribution width (RDW) is correlated with the severity and prognosis of cardiovascular disease. The target of our study was to assess the relationship between RDW and the prognosis of ischemic cardiomyopathy (ICM) patients undergoing percutaneous coronary intervention (PCI). Methods: The study retrospectively enrolled 1986 ICM patients undergoing PCI. The patients were divided into three groups by RDW tertiles. The primary endpoint was major adverse cardiovascular events (MACE) and the secondary endpoints were each of the components of MACE (all-cause mortality, nonfatal myocardial infarction (MI) and any revascularization). Kaplan–Meier survival analyses were conducted to show the association between RDW and the incidence of adverse outcomes. The independent effect of RDW on adverse outcomes was determined by multivariate Cox proportional hazard regression analysis. In addition, the nonlinear relationship between RDW values and MACE was explored using restricted cubic spline (RCS) analysis. The relationship between RDW and MACE in different subgroups was determined using subgroup analysis. Results: As RDW tertiles increased, the incidences of MACE (Tertile 3 vs. Tertile 1: 42.6 vs. 23.7, p p p p p p p for trend p for trend p for trend < 0.001). In addition, the RCS analysis suggested nonlinear association between RDW values and MACE. The subgroup analysis revealed that elderly patients or patients with angiotensin receptor blockers (ARBs) had a higher risk of MACE with higher RDW. Patients with hypercholesterolemia or without anemia also had a higher risk of MACE. Conclusions: RDW was significantly related to the increased risk of MACE among ICM patients undergoing PCI