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Description

Precise spatial positioning and isolation of mammalian cells is a critical component of many single cell experimental methods and biological engineering applications. Although a variety of cell patterning methods have been demonstrated, many of these methods subject cells to high stress environments, discriminate against certain phenotypes, or are a challenge

Precise spatial positioning and isolation of mammalian cells is a critical component of many single cell experimental methods and biological engineering applications. Although a variety of cell patterning methods have been demonstrated, many of these methods subject cells to high stress environments, discriminate against certain phenotypes, or are a challenge to implement. Here, we demonstrate a rapid, simple, indiscriminate, and minimally perturbing cell patterning method using a laser fabricated polymer stencil. The stencil fabrication process requires no stencil-substrate alignment, and is readily adaptable to various substrate geometries and experiments.

ContributorsMessner, Jacob J. (Author) / Glenn, Honor (Author) / Meldrum, Deirdre (Author) / Biodesign Institute (Contributor)
Created2017-12-19
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Description

Quantitative three-dimensional (3D) computed tomography (CT) imaging of living single cells enables orientation-independent morphometric analysis of the intricacies of cellular physiology. Since its invention, x-ray CT has become indispensable in the clinic for diagnostic and prognostic purposes due to its quantitative absorption-based imaging in true 3D that allows objects of

Quantitative three-dimensional (3D) computed tomography (CT) imaging of living single cells enables orientation-independent morphometric analysis of the intricacies of cellular physiology. Since its invention, x-ray CT has become indispensable in the clinic for diagnostic and prognostic purposes due to its quantitative absorption-based imaging in true 3D that allows objects of interest to be viewed and measured from any orientation. However, x-ray CT has not been useful at the level of single cells because there is insufficient contrast to form an image. Recently, optical CT has been developed successfully for fixed cells, but this technology called Cell-CT is incompatible with live-cell imaging due to the use of stains, such as hematoxylin, that are not compatible with cell viability. We present a novel development of optical CT for quantitative, multispectral functional 4D (three spatial + one spectral dimension) imaging of living single cells. The method applied to immune system cells offers truly isotropic 3D spatial resolution and enables time-resolved imaging studies of cells suspended in aqueous medium. Using live-cell optical CT, we found a heterogeneous response to mitochondrial fission inhibition in mouse macrophages and differential basal remodeling of small (0.1 to 1 fl) and large (1 to 20 fl) nuclear and mitochondrial structures on a 20- to 30-s time scale in human myelogenous leukemia cells. Because of its robust 3D measurement capabilities, live-cell optical CT represents a powerful new tool in the biomedical research field.

ContributorsKelbauskas, Laimonas (Author) / Shetty, Rishabh Manoj (Author) / Cao, Bin (Author) / Wang, Kuo-Chen (Author) / Smith, Dean (Author) / Wang, Hong (Author) / Chao, Shi-Hui (Author) / Gangaraju, Sandhya (Author) / Ashcroft, Brian (Author) / Kritzer, Margaret (Author) / Glenn, Honor (Author) / Johnson, Roger (Author) / Meldrum, Deirdre (Author) / Biodesign Institute (Contributor)
Created2017-12-06
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Description

Sustainable fisheries management is key to restoring and maintaining ecological function and benefits to people, but it requires accurate information about patterns of resource use, particularly fishing pressure. In most coral reef fisheries and other data-poor contexts, obtaining such information is challenging and remains an impediment to effective management. We

Sustainable fisheries management is key to restoring and maintaining ecological function and benefits to people, but it requires accurate information about patterns of resource use, particularly fishing pressure. In most coral reef fisheries and other data-poor contexts, obtaining such information is challenging and remains an impediment to effective management. We developed the most comprehensive regional view of shore-based fishing effort and catch published to date, to show detailed fishing patterns from across the main Hawaiian Islands (MHI). We reveal these regional patterns through fisher “creel” surveys conducted by local communities, state agencies, academics, and/or environmental organizations, at 18 sites, comprising >10,000 h of monitoring across a range of habitats and human influences throughout the MHI. All creel surveys included in this study except for one were previously published in some form (peer-reviewed articles or gray literature reports). Here, we synthesize these studies to document spatial patterns in nearshore fisheries catch, effort, catch rates (i.e., catch-per-unit-effort (CPUE)), and catch disposition (i.e., use of fish after catch is landed). This effort provides for a description of general regional patterns based on these location-specific studies. Line fishing was by far the dominant gear type employed. The most efficient gear (i.e., highest CPUE) was spear (0.64 kg h-1), followed closely by net (0.61 kg h-1), with CPUE for line (0.16 kg h-1) substantially lower than the other two methods. Creel surveys also documented illegal fishing activity across the studied locations, although these activities were not consistent across sites. Overall, most of the catch was not sold, but rather retained for home consumption or given away to extended family, which suggests that cultural practices and food security may be stronger drivers of fishing effort than commercial exploitation for coral reef fisheries in Hawai‘i. Increased monitoring of spatial patterns in nearshore fisheries can inform targeted management, and can help communities develop a more informed understanding of the drivers of marine resource harvest and the state of the resources, in order to maintain these fisheries for food security, cultural practices, and ecological value.

Created2017-12-04
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Description

With the successful development of organic/polymeric light emitting diodes, many organic and polymeric fluorophores with high quantum efficiencies and optical stability were synthesized. However, most of these materials which have excellent optical properties are insoluble in water, limiting their applications in biological fields. Herein, we used micelles formed from an

With the successful development of organic/polymeric light emitting diodes, many organic and polymeric fluorophores with high quantum efficiencies and optical stability were synthesized. However, most of these materials which have excellent optical properties are insoluble in water, limiting their applications in biological fields. Herein, we used micelles formed from an amino-group-containing poly(ε-caprolactone)-block-poly(ethylene glycol) (PCL-b-PEG-NH2) to incorporate a hydrophobic blue emitter oligofluorene (OF) to enable its application in biological conditions. Although OF is completely insoluble in water, it was successfully transferred into aqueous solutions with a good retention of its photophysical properties. OF exhibited a high quantum efficiency of 0.84 in a typical organic solvent of tetrahydrofuran (THF). In addition, OF also showed a good quantum efficiency of 0.46 after being encapsulated into micelles. Two cells lines, human glioblastoma (U87MG) and esophagus premalignant (CP-A), were used to study the cellular internalization of the OF incorporated micelles. Results showed that the hydrophobic OF was located in the cytoplasm, which was confirmed by co-staining the cells with nucleic acid specific SYTO 9, lysosome specific LysoTracker Red®, and mitochondria specific MitoTracker Red. MTT assay indicated non-toxicity of the OF-incorporated micelles. This study will broaden the application of hydrophobic functional organic compounds, oligomers, and polymers with good optical properties to enable their applications in biological research fields.

ContributorsSu, Fengyu (Author) / Alam, Ruhaniyah (Author) / Mei, Qian (Author) / Tian, Yanqing (Author) / Meldrum, Deirdre (Author) / Biodesign Institute (Contributor)
Created2011-09-06
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Description

Three studies examined the symbolic and self-presentational meaning of low-water-use residential landscaping in a desert city in the southwestern United States. We hypothesized that owners' water-intensive or water-conserving landscape choices would be seen to convey very different characteristics. Data indicated that these two types of residential landscapes led to substantially

Three studies examined the symbolic and self-presentational meaning of low-water-use residential landscaping in a desert city in the southwestern United States. We hypothesized that owners' water-intensive or water-conserving landscape choices would be seen to convey very different characteristics. Data indicated that these two types of residential landscapes led to substantially different attributions about homeowners and also that potential homeowners could use landscapes to convey an array of characteristics to a social audience. In general, water-intensive landscapes led to more positive attributions than did water-conserving landscapes. The results support the idea that landscaping choice may be guided by self-presentational considerations, and that such considerations might influence the adoption of high- or low-water-use landscapes.

ContributorsNeel, Rebecca (Author) / Sadalla, Edward (Author) / Berlin, Anna (Author) / Ledlow, Susan (Author) / Neufeld, Samantha (Author) / Julie Ann Wrigley Global Institute of Sustainability (Contributor)
Created2014-12-01
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Description

Hydrophobic platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorophenyl)-porphyrin (PtTFPP) was physically incorporated into micelles formed from poly(ε-caprolactone)-block-poly(ethylene glycol) to enable the application of PtTFPP in aqueous solution. Micelles were characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM) to show an average diameter of about 140 nm. PtTFPP showed higher quantum efficiency in micellar

Hydrophobic platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorophenyl)-porphyrin (PtTFPP) was physically incorporated into micelles formed from poly(ε-caprolactone)-block-poly(ethylene glycol) to enable the application of PtTFPP in aqueous solution. Micelles were characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM) to show an average diameter of about 140 nm. PtTFPP showed higher quantum efficiency in micellar solution than in tetrahydrofuran (THF) and dichloromethane (CH2Cl2). PtTFPP in micelles also exhibited higher photostability than that of PtTFPP suspended in water. PtTFPP in micelles exhibited good oxygen sensitivity and response time. This study provided an efficient approach to enable the application of hydrophobic oxygen sensors in a biological environment.

ContributorsSu, Fengyu (Author) / Alam, Ruhaniyah (Author) / Mei, Qian (Author) / Tian, Yanqing (Author) / Youngbull, Cody (Author) / Johnson, Roger (Author) / Meldrum, Deirdre (Author) / Biodesign Institute (Contributor)
Created2012-03-22
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Description

Conservation of marine resources is critical to the wellbeing of human communities. Coastal artisanal fishing communities are particularly reliant on marine resources for food and for their livelihoods. Management actions aimed at marine conservation may lead to unanticipated changes in human behavior that influence the ability of conservation programs to

Conservation of marine resources is critical to the wellbeing of human communities. Coastal artisanal fishing communities are particularly reliant on marine resources for food and for their livelihoods. Management actions aimed at marine conservation may lead to unanticipated changes in human behavior that influence the ability of conservation programs to achieve their goals. We examine how marine conservation strategies may impact labor decisions that influence both the ecosystem and human livelihoods using simulation modeling. We consider two conservation strategies in the model: direct action through fisheries regulation enforcement, and indirect action through land conservation. Our results indicate that both strategies can increase the abundance of fish, and thus contribute to the maintenance of marine resources. However, our results also show that marine fisheries enforcement may negatively impact the livelihoods of human communities. Land conservation, on the other hand, potentially enhances the livelihood of the human populations. Thus, depending on management objectives, indirect or a combination of direct and indirect conservation strategies may be effective at achieving conservation and sustainability goals. These results highlight the importance of accounting for changes in human behavior resulting from management actions in conservation and management.

ContributorsHughes, Zachary (Author) / Fenichel, Eli (Author) / Gerber, Leah (Author) / Julie Ann Wrigley Global Institute of Sustainability (Contributor)
Created2011-08-24
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Description

Background: The use of culture-independent nucleic acid techniques, such as ribosomal RNA gene cloning library analysis, has unveiled the tremendous microbial diversity that exists in natural environments. In sharp contrast to this great achievement is the current difficulty in cultivating the majority of bacterial species or phylotypes revealed by molecular approaches.

Background: The use of culture-independent nucleic acid techniques, such as ribosomal RNA gene cloning library analysis, has unveiled the tremendous microbial diversity that exists in natural environments. In sharp contrast to this great achievement is the current difficulty in cultivating the majority of bacterial species or phylotypes revealed by molecular approaches. Although recent new technologies such as metagenomics and metatranscriptomics can provide more functionality information about the microbial communities, it is still important to develop the capacity to isolate and cultivate individual microbial species or strains in order to gain a better understanding of microbial physiology and to apply isolates for various biotechnological applications.

Results: We have developed a new system to cultivate bacteria in an array of droplets. The key component of the system is the microbe observation and cultivation array (MOCA), which consists of a Petri dish that contains an array of droplets as cultivation chambers. MOCA exploits the dominance of surface tension in small amounts of liquid to spontaneously trap cells in well-defined droplets on hydrophilic patterns. During cultivation, the growth of the bacterial cells across the droplet array can be monitored using an automated microscope, which can produce a real-time record of the growth. When bacterial cells grow to a visible microcolony level in the system, they can be transferred using a micropipette for further cultivation or analysis.

Conclusions: MOCA is a flexible system that is easy to set up, and provides the sensitivity to monitor growth of single bacterial cells. It is a cost-efficient technical platform for bioassay screening and for cultivation and isolation of bacteria from natural environments.

ContributorsGao, Weimin (Author) / Navarroli, Dena (Author) / Naimark, Jared (Author) / Zhang, Weiwen (Author) / Chao, Shih-hui (Author) / Meldrum, Deirdre (Author) / Biodesign Institute (Contributor)
Created2013-01-09
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Description

Smart growth policy and planning have tended to emphasize urban centers and regions, yet rural communities can also be important sites of innovation. Recent work demonstrated that these communities had surprisingly high levels of current and potential nonmotorized travel. Legislation in California mandates reductions in greenhouse gas emissions across all

Smart growth policy and planning have tended to emphasize urban centers and regions, yet rural communities can also be important sites of innovation. Recent work demonstrated that these communities had surprisingly high levels of current and potential nonmotorized travel. Legislation in California mandates reductions in greenhouse gas emissions across all of the state's metropolitan planning organization (MPO) regions, including the heavily rural San Joaquin Valley. Advocates for rural communities are finding common cause with more traditional environmental organizations around the vision of investing in and enhancing extant rural places as an alternative to leapfrog patterns of urban and suburban sprawl. Because of existing patterns of extreme disparity and legion underserved unincorporated communities, analyses that can help integrate social equity within regional planning are needed to serve and empower rural residents. This paper presents the results of several new analyses of the social equity dimensions of regional transportation plans in the San Joaquin Valley. Activity-based travel model data were used to analyze equity, with a particular focus placed on eight disadvantaged unincorporated communities identified by community advocates to be important demonstration sites. The investigators showed how improvements to traditional equity analysis could enhance the consideration of equity in the planning process and compared the results developed by innovative techniques with those obtained by use of their traditional counterparts. The methods outlined here can make substantial contributions to reduce disparities in rural communities, which would likely be overlooked in typical regional equity analyses because of their small size, and offer lessons for MPOs serving rural areas across the country.

Created2013-11-30
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Description

During the last 40 years evidence from systematic case study analysis and behavioral experiments have provided a comprehensive perspective on how communities can manage common resources in a sustainable way. The conventional theory based on selfish rational actors cannot explain empirical observations. A more comprehensive theoretical framework of human behavior

During the last 40 years evidence from systematic case study analysis and behavioral experiments have provided a comprehensive perspective on how communities can manage common resources in a sustainable way. The conventional theory based on selfish rational actors cannot explain empirical observations. A more comprehensive theoretical framework of human behavior is emerging that include concepts such as trust, conditional cooperation, other-regarding preferences, social norms, and reputation. The new behavioral perspective also demonstrates that behavioral responses depend on social and biophysical context.

Created2015-02-01