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- Barbe, Laurent, et al.
(författare)
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Toward a confocal subcellular atlas of the human proteome
- 2008
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Ingår i: Molecular and cellular proteomics. - 1535-9476 .- 1535-9484. ; 7:3, s. 499-508
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Tidskriftsartikel (refereegranskat)abstract
- Information on protein localization on the subcellular level is important to map and characterize the proteome and to better understand cellular functions of proteins. Here we report on a pilot study of 466 proteins in three human cell lines aimed to allow large scale confocal microscopy analysis using protein-specific antibodies. Approximately 3000 high resolution images were generated, and more than 80% of the analyzed proteins could be classified in one or multiple subcellular compartment(s). The localizations of the proteins showed, in many cases, good agreement with the Gene Ontology localization prediction model. This is the first large scale antibody-based study to localize proteins into subcellular compartments using antibodies and confocal microscopy. The results suggest that this approach might be a valuable tool in conjunction with predictive models for protein localization.
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| 5. |
- Cantoni, Federico, et al.
(författare)
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A carrier with an integrated perfusion and heating systems for long-term imaging of microfluidic chips
- 2021
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Konferensbidrag (refereegranskat)abstract
- Microfluidic chips offer many benefits for cell studies, including an accurate spatial and temporal control over the growth conditions1. Despite the large expansion of microfluidics in biological applications2, there have been only a few developments of devices to simplify microfluidic chip handling and imaging. Here, we present a carrier of well-plate format with integrated cell media recirculation and heating systems to provide a stable environment for the cell culture during the imaging outside the incubator. Moreover, the absence of external tubing reduces the risk of contamination and bubble formation during the carrier transfers and reagent injections enabling long-term experiment monitoring. Our system was validated by repeatedly (day 1, 3, 7 and 10) taking the cultured mouse endothelial cells (bEnd.3) out of the incubator for imaging.
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| 6. |
- Cantoni, Federico, et al.
(författare)
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A microfluidic chip carrier including temperature control and perfusion system for long-term cell imaging
- 2021
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Ingår i: HardwareX. - : Elsevier. - 2468-0672. ; 10, s. e00245-
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Tidskriftsartikel (refereegranskat)abstract
- Microfluidic devices are widely used for biomedical applications but there is still a lack of affordable, reliable and user-friendly systems for transferring microfluidic chips from an incubator to a microscope while maintaining physiological conditions when performing microscopy. The presented carrier represents a cost-effective option for sustaining environmental conditions of microfluidic chips in combination with minimizing the device manipulation required for reagent injection, media exchange or sample collection. The carrier, which has the outer dimension of a standard well plate size, contains an integrated perfusion system that can recirculate the media using piezo pumps, operated in either continuous or intermittent modes (50–1000 µl/min). Furthermore, a film resistive heater made from 37 µm-thick copper wires, including temperature feedback control, was used to maintain the microfluidic chip temperature at 37 °C when outside the incubator. The heater characterisation showed a uniform temperature distribution along the chip channel for perfusion flow rates up to 10 µl/min. To demonstrate the feasibility of our platform for long term cell culture monitoring, mouse brain endothelial cells (bEnd.3) were repeatedly monitored for a period of 10 days, demonstrating a system with both the versatility and the potential for long imaging in microphysiological system cell cultures.
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| 7. |
- Cantoni, Federico, et al.
(författare)
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A perfusable multi-hydrogel vasculature on-chip engineered by 2-photon 3D printing and scaffold molding to improve microfabrication fidelity in hydrogels
- 2024
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Ingår i: Advanced Materials Technologies. - : John Wiley & Sons. - 2365-709X. ; 9:4
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Tidskriftsartikel (refereegranskat)abstract
- Engineering vasculature networks in physiologically relevant hydrogelsrepresents a challenge in terms of both fabrication, due to the cell–bioinkinteractions, as well as the subsequent hydrogel-device interfacing. Here, anew cell-friendly fabrication strategy is presented to realize perfusablemulti-hydrogel vasculature models supporting co-culture integrated in amicrofluidic chip. The system comprises two different hydrogels to specificallysupport the growth and proliferation of two different cell types selected for thevessel model. First, the channels are printed in a gelatin-based ink bytwo-photon polymerization (2PP) inside the microfluidic device. Then, ahuman lung fibroblast-laden fibrin hydrogel is injected to surround the printednetwork. Finally, human endothelial cells are seeded inside the printedchannels. The printing parameters and fibrin composition are optimized toreduce hydrogel swelling and ensure a stable model that can be perfused withcell media. Fabricating the hydrogel structure in two steps ensures that nocells are exposed to cytotoxic fabrication processes, while still obtaining highfidelity printing. In this work, the possibility to guide the endothelial cellinvasion through the 3D printed scaffold and perfusion of the co-culturemodel for 10 days is successfully demonstrated on a custom-made perfusionsystem.
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| 10. |
- Cantoni, Federico, et al.
(författare)
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Round-robin testing of commercial two-photon polymerization 3D printers
- 2023
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Ingår i: Additive Manufacturing. - : Elsevier. - 2214-8604 .- 2214-7810. ; 76
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Tidskriftsartikel (refereegranskat)abstract
- Since its introduction in the 1980s, 3D printing has advanced as a versatile and reliable tool with applications in different fields. Among the available 3D printing techniques, two-photon polymerization is regarded as one of the most promising technologies for microscale printing due to its ability to combine a high printing fidelity down to submicron scale with free-form structure design. Recently, the technology has been enhanced through the implementation of faster laser scanning strategies, as well as the development of new photoresists. This paves the way for a wide range of applications, which has resulted in an increasing number of available commercial systems. This work aims to provide an overview of the technology capability by comparing three commercial systems in a round-robin test. To cover a wide range of applications, six test structures with distinct features were designed, covering various aspects of interest, from single material objects with sub-micron feature sizes up to multi-material millimeter-sized objects. Application-specific structures were printed to evaluate surface roughness and the stitching capability of the printers. Moreover, the ability to generate free-hanging structures and complex surfaces required for cell scaffolds and microfluidic platform fabrication was quantitatively investigated. Finally, the influence of the numerical aperture of the fabrication objective on the printing quality was assessed. All three printers successfully fabricated samples comprising various three-dimensional features and achieved submicron resolution and feature sizes, demonstrating the versatility and precision of two-photon polymerization direct laser writing. Our study will facilitate the understanding of the technology maturity level, while highlighting specific aspects that characterize each of the investigated systems.
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