| 1. |
- Blom, M., et al.
(författare)
-
RhoD is a Golgi component with a role in anterograde protein transport from the ER to the plasma membrane
- 2015
-
Ingår i: Experimental Cell Research. - : Elsevier BV. - 0014-4827 .- 1090-2422. ; 333:2, s. 208-219
-
Tidskriftsartikel (refereegranskat)abstract
- RhoD is a member of the Rho GTPase family and it coordinates actin dynamics and membrane trafficking. Activation of RhoD results in formation of filopodia, dissolution of stress fibers, and the subsequent formation of short actin bundles. In addition, RhoD localizes to early endosomes and recycling endosomes, and has a regulatory role in endosome trafficking. In this study, we report on a function of RhoD in the regulation of Golgi homeostasis. We show that manipulation of protein and activation levels of RhoD, as well as of its binding partner WHAMM, result in derailed localization of Golgi stacks. Moreover, vesicle trafficking from the endoplasmic reticulum to the plasma membrane via the Golgi apparatus measured by the VSV-G protein is severely hampered by manipulation of RhoD or WHAMM. In summary, our studies demonstrate a novel role for this member of the Rho GTPases in the regulation of Golgi function.
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| 2. |
- Kaucka, Marketa, et al.
(författare)
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Analysis of neural crest-derived clones reveals novel aspects of facial development
- 2016
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Ingår i: Science Advances. - : American Association for the Advancement of Science. - 2375-2548. ; 2:8
-
Tidskriftsartikel (refereegranskat)abstract
- Cranial neural crest cells populate the future facial region and produce ectomesenchyme-derived tissues, such as cartilage, bone, dermis, smooth muscle, adipocytes, and many others. However, the contribution of individual neural crest cells to certain facial locations and the general spatial clonal organization of the ectomesenchyme have not been determined. We investigated how neural crest cells give rise to clonally organized ectomesenchyme and how this early ectomesenchyme behaves during the developmental processes that shape the face. Using a combination of mouse and zebrafish models, we analyzed individual migration, cell crowd movement, oriented cell division, clonal spatial overlapping, and multilineage differentiation. The early face appears to be built from multiple spatially defined overlapping ectomesenchymal clones. During early face development, these clones remain oligopotent and generate various tissues in a given location. By combining clonal analysis, computer simulations, mouse mutants, and live imaging, we show that facial shaping results from an array of local cellular activities in the ectomesenchyme. These activities mostly involve oriented divisions and crowd movements of cells during morphogenetic events. Cellular behavior that can be recognized as individual cell migration is very limited and short-ranged and likely results from cellular mixing due to the proliferation activity of the tissue. These cellular mechanisms resemble the strategy behind limb bud morphogenesis, suggesting the possibility of common principles and deep homology between facial and limb outgrowth.
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| 3. |
- Kaukua, Nina, et al.
(författare)
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Glial origin of mesenchymal stem cells in a tooth model system
- 2014
-
Ingår i: Nature. - : Springer Science and Business Media LLC. - 0028-0836 .- 1476-4687. ; 513:7519, s. 551-554
-
Tidskriftsartikel (refereegranskat)abstract
- Mesenchymal stem cells occupy niches in stromal tissues where they provide sources of cells for specialized mesenchymal derivatives during growth and repair(1). The origins of mesenchymal stem cells have been the subject of considerable discussion, and current consensus holds that perivascular cells form mesenchymal stem cells in most tissues. The continuously growing mouse incisor tooth offers an excellent model to address the origin of mesenchymal stem cells. These stem cells dwell in a niche at the tooth apex where they produce a variety of differentiated derivatives. Cells constituting the tooth are mostly derived from two embryonic sources: neural crest ectomesenchyme and ectodermal epithelium(2). It has been thought for decades that the dental mesenchymal stem cells(3) giving rise to pulp cells and odontoblasts derive from neural crest cells after their migration in the early head and formation of ectomesenchymal tissue(4,5). Here we show that a significant population of mesenchymal stem cells during development, self-renewal and repair of a tooth are derived from peripheral nerve-associated glia. Glial cells generate multipotent mesenchymal stem cells that produce pulp cells and odontoblasts. By combining a clonal colour-coding technique(6) with tracing of peripheral glia, we provide new insights into the dynamics of tooth organogenesis and growth.
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| 4. |
- Reuss, Matthias, et al.
(författare)
-
Measuring true localization accuracy in super resolution microscopy with DNA-origami nanostructures
- 2017
-
Ingår i: New Journal of Physics. - : Institute of Physics Publishing (IOPP). - 1367-2630. ; 19:2
-
Tidskriftsartikel (refereegranskat)abstract
- A common method to assess the performance of (super resolution) microscopes is to use the localization precision of emitters as an estimate for the achieved resolution. Naturally, this is widely used in super resolution methods based on single molecule stochastic switching. This concept suffers from the fact that it is hard to calibrate measures against a real sample (a phantom), because true absolute positions of emitters are almost always unknown. For this reason, resolution estimates are potentially biased in an image since one is blind to true position accuracy, i.e. deviation in position measurement from true positions. We have solved this issue by imaging nanorods fabricated with DNA-origami. The nanorods used are designed to have emitters attached at each end in a well-defined and highly conserved distance. These structures are widely used to gauge localization precision. Here, we additionally determined the true achievable localization accuracy and compared this figure of merit to localization precision values for two common super resolution microscope methods STED and STORM.
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| 5. |
- Stenudd, Moa, et al.
(författare)
-
Identification of a discrete subpopulation of spinal cord ependymal cells with neural stem cell properties
- 2022
-
Ingår i: Cell Reports. - : CELL PRESS. - 2211-1247. ; 38:9
-
Tidskriftsartikel (refereegranskat)abstract
- Spinal cord ependymal cells display neural stem cell properties in vitro and generate scar-forming astrocytes and remyelinating oligodendrocytes after injury. We report that ependymal cells are functionally heterogeneous and identify a small subpopulation (8% of ependymal cells and 0.1% of all cells in a spinal cord segment), which we denote ependymal A (EpA) cells, that accounts for the in vitro stem cell potential in the adult spinal cord. After spinal cord injury, EpA cells undergo self-renewing cell division as they give rise to differentiated progeny. Single-cell transcriptome analysis revealed a loss of ependymal cell gene expression programs as EpA cells gained signaling entropy and dedifferentiated to a stem-cell-like transcriptional state after an injury. We conclude that EpA cells are highly differentiated cells that can revert to a stem cell state and constitute a therapeutic target for spinal cord repair.
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| 6. |
- Abrahamsson, S., et al.
(författare)
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Multifocus structured illumination microscopy for fast volumetric super-resolution imaging
- 2017
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Ingår i: Biomedical Optics Express. - : OSA - The Optical Society. - 2156-7085. ; 8:9, s. 4135-4140
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Tidskriftsartikel (refereegranskat)abstract
- We here report for the first time the synergistic implementation of structured illumination microscopy (SIM) and multifocus microscopy (MFM). This imaging modality is designed to alleviate the problem of insufficient volumetric acquisition speed in superresolution biological imaging. SIM is a wide-field super-resolution technique that allows imaging with visible light beyond the classical diffraction limit. Employing multifocus diffractive optics we obtain simultaneous wide-field 3D imaging capability in the SIM acquisition sequence, improving volumetric acquisition speed by an order of magnitude. Imaging performance is demonstrated on biological specimens.
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| 7. |
- Agostinho, Ana, et al.
(författare)
-
High density of REC8 constrains sister chromatid axes and prevents illegitimate synaptonemal complex formation
- 2016
-
Ingår i: EMBO Reports. - Stockholm : Wiley-VCH Verlagsgesellschaft. - 1469-221X .- 1469-3178. ; 17:6, s. 901-913
-
Tidskriftsartikel (refereegranskat)abstract
- During meiosis, cohesin complexes mediate sister chromatid cohesion (SCC), synaptonemal complex (SC) assembly and synapsis. Here, using super-resolution microscopy, we imaged sister chromatid axes in mouse meiocytes that have normal or reduced levels of cohesin complexes, assessing the relationship between localization of cohesin complexes, SCC and SC formation. We show that REC8 foci are separated from each other by a distance smaller than 15% of the total chromosome axis length in wild-type meiocytes. Reduced levels of cohesin complexes result in a local separation of sister chromatid axial elements (LSAEs), as well as illegitimate SC formation at these sites. REC8 but not RAD21 or RAD21L cohesin complexes flank sites of LSAEs, whereas RAD21 and RAD21L appear predominantly along the separated sister-chromatid axes. Based on these observations and a quantitative distribution analysis of REC8 along sister chromatid axes, we propose that the high density of randomly distributed REC8 cohesin complexes promotes SCC and prevents illegitimate SC formation.
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| 8. |
- Agostinho, A., et al.
(författare)
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Sexual dimorphism in the width of the mouse synaptonemal complex
- 2018
-
Ingår i: Journal of Cell Science. - : Company of Biologists Ltd. - 0021-9533 .- 1477-9137. ; 131:5
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Tidskriftsartikel (refereegranskat)abstract
- Sexual dimorphism has been used to describe morphological differences between the sexes, but can be extended to any biologically related process that varies between males and females. The synaptonemal complex (SC) is a tripartite structure that connects homologous chromosomes in meiosis. Here, aided by superresolution microscopy techniques, we show that the SC is subject to sexual dimorphism, in mouse germ cells. We have identified a significantly narrower SC in oocytes and have established that this difference does not arise from a different organization of the lateral elements nor from a different isoform of transverse filament protein SYCP1. Instead, we provide evidence for the existence of a narrower central element and a different integration site for the C-termini of SYCP1, in females. In addition to these female-specific features, we speculate that post-translation modifications affecting the SYCP1 coiled-coil region could render a more compact conformation, thus contributing to the narrower SC observed in females.
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| 9. |
- Akkuratov, Evgeny E., et al.
(författare)
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Ouabain Modulates the Functional Interaction Between Na,K-ATPase and NMDA Receptor.
- 2020
-
Ingår i: Molecular Neurobiology. - : Springer Science and Business Media LLC. - 0893-7648 .- 1559-1182. ; 57:10, s. 4018-4030
-
Tidskriftsartikel (refereegranskat)abstract
- The N-methyl-D-aspartate (NMDA) receptor plays an essential role in glutamatergic transmission and synaptic plasticity and researchers are seeking for different modulators of NMDA receptor function. One possible mechanism for its regulation could be through adjacent membrane proteins. NMDA receptors coprecipitate with Na,K-ATPase, indicating a potential interaction of these two proteins. Ouabain, a mammalian cardiotonic steroid that specifically binds to Na,K-ATPase and affects its conformation, can protect from some toxic effects of NMDA receptor activation. Here we have examined whether NMDA receptor activity and downstream effects can be modulated by physiological ouabain concentrations. The spatial colocalization between NMDA receptors and the Na,K-ATPase catalytic subunits on dendrites of cultured rat hippocampal neurons was analyzed with super-resolution dSTORM microscopy. The functional interaction was analyzed with calcium imaging of single hippocampal neurons exposed to 10 μM NMDA in presence and absence of ouabain and by determination of the ouabain effect on NMDA receptor-dependent long-term potentiation. We show that NMDA receptors and the Na,K-ATPase catalytic subunits alpha1 and alpha3 exist in same protein complex and that ouabain in nanomolar concentration consistently reduces the calcium response to NMDA. Downregulation of the NMDA response is not associated with internalization of the receptor or with alterations in its state of Src phosphorylation. Ouabain in nanomolar concentration elicits a long-term potentiation response. Our findings suggest that ouabain binding to a fraction of Na,K-ATPase molecules that cluster with the NMDA receptors will, via a conformational effect on the NMDA receptors, cause moderate but consistent reduction of NMDA receptor response at synaptic activation.
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| 10. |
- Allesøe, Rosa Lundbye, et al.
(författare)
-
Discovery of drug–omics associations in type 2 diabetes with generative deep-learning models
- 2023
-
Ingår i: Nature Biotechnology. - : Springer Nature. - 1087-0156 .- 1546-1696. ; 41:3, s. 399-408
-
Tidskriftsartikel (refereegranskat)abstract
- The application of multiple omics technologies in biomedical cohorts has the potential to reveal patient-level disease characteristics and individualized response to treatment. However, the scale and heterogeneous nature of multi-modal data makes integration and inference a non-trivial task. We developed a deep-learning-based framework, multi-omics variational autoencoders (MOVE), to integrate such data and applied it to a cohort of 789 people with newly diagnosed type 2 diabetes with deep multi-omics phenotyping from the DIRECT consortium. Using in silico perturbations, we identified drug–omics associations across the multi-modal datasets for the 20 most prevalent drugs given to people with type 2 diabetes with substantially higher sensitivity than univariate statistical tests. From these, we among others, identified novel associations between metformin and the gut microbiota as well as opposite molecular responses for the two statins, simvastatin and atorvastatin. We used the associations to quantify drug–drug similarities, assess the degree of polypharmacy and conclude that drug effects are distributed across the multi-omics modalities.
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| 11. |
-
Att välja trä
- 2020. - 10
-
Samlingsverk (redaktörskap) (populärvet., debatt m.m.)
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| 12. |
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| 13. |
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| 14. |
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| 15. |
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| 16. |
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| 17. |
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| 18. |
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| 19. |
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| 20. |
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| 21. |
- Bennemo, Mia, et al.
(författare)
-
A chromatographic method for determination of supercoiled plasmid DNA concentration in complex solutions.
- 2009
-
Ingår i: Journal of chromatography. B. - : Elsevier BV. - 1570-0232 .- 1873-376X. ; 877:24, s. 2530-6
-
Tidskriftsartikel (refereegranskat)abstract
- A method for determination of the plasmid DNA concentration with subsequent analysis of the ratio supercoiled to open circular form is presented. The method is suitable for samples from all steps of the manufacturing process, from fermentation to final product. The analysis consists of size exclusion chromatography, followed by analytical thiophilic aromatic chromatography. In the first step, the plasmid DNA concentration is determined by group separation, including removal of RNA and other impurities, within less than 2 min. The limit of detection and quantification was 0.28 and 0.83 microg/ml, respectively. The precision of the method is high, providing a coefficient of variation as low as below 2%. In the second step, the ratio of open circular to supercoiled plasmid DNA is determined following separation of the two plasmid DNA isoforms with a linear salt gradient. The precision of the second step was evaluated using serial injections of aliquots of a sample stock solution. In comparison with the two most commonly used methods, the developed analysis was found to be significantly more accurate than agarose gel electrophoresis and equivalent to capillary gel electrophoresis. The combined methods for quantification and control of homogeneity of plasmid DNA presented here enable reliable and precise analysis at all steps of the manufacturing process.
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| 22. |
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| 23. |
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| 24. |
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| 25. |
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| 26. |
- Berg, Sören, et al.
(författare)
-
Process modelling of reactive sputtering
- 1989
-
Ingår i: Journal of Vacuum Science & Technology. A. Vacuum, Surfaces, and Films. - 0734-2101 .- 1520-8559. ; 7:3, s. 1225-1229
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Tidskriftsartikel (refereegranskat)
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| 27. |
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| 28. |
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| 29. |
- Bernhem, Kristoffer, et al.
(författare)
-
Quantification of endogenous and exogenous protein expressions of Na,K-ATPase with super-resolution PALM/STORM imaging
-
Annan publikation (populärvet., debatt m.m.)abstract
- Transient transfection of fluorescent fusion proteins is a key enabling technology in fluorescent microscopy to spatio-temporally map cellular protein distributions. Transient transfection of proteins may however bypass normal regulation of expression, leading to overexpression artefacts like misallocations and excess amounts. In this study we investigate the ability to quantitatively monitor endogenous and exogenous protein expression competition on the single molecule level. Through incorporation of an N-terminal hemagglutinin (HA) epitope to amMaple3 fused Na,K-ATPase (α1 isoform), using PALM and STORM imaging we investigatethe increase in plasma membrane density at the cost of competitive expression. Quantification of plasma membrane protein density revealed a time dependent increase over time of totalprotein content. Results show that plasma membrane densities increased by more than 60%,comparing 17h and 41h transfection times, whilst endogenous levels were simultaneously reduced by 20 %.
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| 30. |
- Bernhem, Kristoffer, et al.
(författare)
-
Quantification of endogenous and exogenous protein expressions of Na,K-ATPase with super-resolution PALM/STORM imaging
- 2018
-
Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203.
-
Tidskriftsartikel (refereegranskat)abstract
- Transient transfection of fluorescent fusion proteins is a key enabling technology in fluorescent microscopy to spatio-temporally map cellular protein distributions. Transient transfection of proteins may however bypass normal regulation of expression, leading to overexpression artefacts like misallocations and excess amounts. In this study we investigate the use of STORM and PALM microscopy to quantitatively monitor endogenous and exogenous protein expression. Through incorporation of an N-terminal hemagglutinin epitope to a mMaple3 fused Na,K-ATPase (α1 isoform), we analyze the spatial and quantitative changes of plasma membrane Na,K-ATPase localization during competitive transient expression. Quantification of plasma membrane protein density revealed a time dependent increase of Na,K-ATPase, but no increase in size of protein clusters. Results show that after 41h transfection, the total plasma membrane density of Na,K-ATPase increased by 63% while the endogenous contribution was reduced by 16%.
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| 31. |
- Bernhem, Kristoffer, et al.
(författare)
-
Super-resolution microscopy reveals that Na+/K+-ATPase signaling protects against glucose-induced apoptosis by deactivating Bad
- 2021
-
Ingår i: Cell Death and Disease. - : Springer Nature. - 2041-4889. ; 12:8
-
Tidskriftsartikel (refereegranskat)abstract
- Activation of the apoptotic pathway is a major cause of progressive loss of function in chronic diseases such as neurodegenerative and diabetic kidney diseases. There is an unmet need for an anti-apoptotic drug that acts in the early stage of the apoptotic process. The multifunctional protein Na+,K+-ATPase has, in addition to its role as a transporter, a signaling function that is activated by its ligand, the cardiotonic steroid ouabain. Several lines of evidence suggest that sub-saturating concentrations of ouabain protect against apoptosis of renal epithelial cells, a common complication and major cause of death in diabetic patients. Here, we induced apoptosis in primary rat renal epithelial cells by exposing them to an elevated glucose concentration (20mM) and visualized the early steps in the apoptotic process using super-resolution microscopy. Treatment with 10nM ouabain interfered with the onset of the apoptotic process by inhibiting the activation of the BH3-only protein Bad and its translocation to mitochondria. This occurred before the pro-apoptotic protein Bax had been recruited to mitochondria. Two ouabain regulated and Akt activating Ca2+/calmodulin-dependent kinases were found to play an essential role in the ouabain anti-apoptotic effect. Our results set the stage for further exploration of ouabain as an anti-apoptotic drug in diabetic kidney disease as well as in other chronic diseases associated with excessive apoptosis.
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| 32. |
- Berntsson, Lars-Olof, et al.
(författare)
-
EAST-ADL 2.0 Specification
- 2008
-
Rapport (refereegranskat)abstract
- This specification of the EAST ADL 2.0 is based on the EAST-ADL developed in the EAST EEA projectand has been further refined and harmonized with on-going modelling appraches in the automotiveindustry. It presents the modeling infrastructure, i.e. how the modeling elements should be represented inthe language and the UML representation. For each package a usage example is provided.The EAST-ADL 2.0 is harmonized with AUTOSAR.The metamodel and UML profile of EAST ADL 2.0 is defined in two steps: A domain (automotive)metamodel is defined, capturing only the domain specific needs of the language, without adding the UML2details. The basic concepts of UML are used for this purpose, such as classes, compositions andassociations. Based on the domain metamodel, a UML2 profile for the domain metamodel is defined,specifying stereotypes with properties and constraints.Comments on the content of this document are welcomed, and should be directed to .Please download the latest available specification and the XMI file ready for use in UML2 tools from the website.
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| 33. |
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| 34. |
- Blom, Hans
(författare)
-
Correlation spectroscopy of single emitters : fundamental studies and applications
- 2001
-
Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Correlation analysis and correlation spectroscopy has eversince the first developments, to characterise light emittingprocesses and biomolecular dynamics, continued to extend itspractical applicability. Today, correlation spectroscopy can beused in life science to study dynamical processes even at thesingle molecule level. Correlation analysis can in one of itsextreme be applied to investigate single photon processes fromsolid-state emitters. This thesis is an account of my studiesof fluorescent emitter related to quantum optics and lifescience. It presents some fundamental results and discussesapplications of emitters like single quantum dots or singledyes attached to biomolecules. The studies were performed bythe means of correlation analysis and correlation spectroscopyon self-made optical setups. One task of this thesis was todevelop fluorescence correlation spectroscopy for ultravioletexcitation and emission. With ultraviolet excitation thenatural intrinsic chromophores of certain nucleotides and aminoacids can be used. No external labelling of biomolecules couldbecome a reality using ultraviolet excitation and emission. Asecond task was to apply correlation spectroscopy to performhigh spatial-resolution flow profiling and trafficking ofsingle dye-labeled biomolecules in microstructured channels.Future transports effects, flow monitoring, flow profiling andprolonged fluorescence detection in artificial microstructuresor in cells, could benefit from this application. An additionaltask was to apply correlation spectroscopy to so-calledmicroarrays for parallel acquisition of dynamical data at thesingle molecule level. Parallel excitation and detection wasachieved with the use of diffractive optical elements andintegrated semiconductor single-photon sensitive detectors. Thecurrent throughput rate in biological diagnostic or screeninganalysis could be increased dramatically with implementation ofthis parallel confocal excitation and detection technique. Yetanother task of this thesis was to investigations single-photongeneration by InAs-semiconductor quantum dots. We show that aquantum dots can be used for single-photon generation ondemand. Besides the single-photon generation in quantum dots,the possibility of two-photon generation, and generation ofentangled photon-pair, has also been investigated
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| 35. |
- Blom, Hans, 1968-
(författare)
-
Correlation spectrosopy of single emitters : fundamental studies and applications related to quantum optics and life science
- 2003
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Correlation analysis and correlation spectroscopy has eversince the first developments, to characterise light emittingprocesses and biomolecular dynamics, continued to extend itspractical applicability. Today, correlation spectroscopy can beused in life science to study dynamical processes even at thesingle molecule level. Correlation analysis can in one of itsextreme be applied to investigate single photon processes fromsolid-state emitters. This thesis is an account of my studiesof fluorescent emitter related to quantum optics and lifescience. It presents some fundamental results and discussesapplicationsof emitters like single quantum dots or singledyes attached to biomolecules. The studies were performed bythe means of correlation analysis and correlation spectroscopyon self-made optical setups. One task of this thesis was todevelop fluorescence correlation spectroscopy for ultravioletexcitation and emission. With ultraviolet excitation thenatural intrinsic chromophores of certain nucleotides and aminoacids can be used. No external labelling of biomolecules couldbecome a reality using ultraviolet excitation and emission. Asecond task was to apply correlation spectroscopy to performhigh spatial-resolution flow profiling and trafficking ofsingle dye-labeled biomolecules in microstructured channels.Future transports effects, flow monitoring, flow profiling andprolonged fluorescence detection in artificial microstructuresor in cells, could benefit from this application. An additionaltask was to apply correlation spectroscopy to so-calledmicroarrays for parallel acquisition of dynamical data at thesingle molecule level. Parallel excitation and detection wasachieved with the use of diffractive optical elements andintegrated semiconductor single-photon sensitive detectors. Thecurrent throughput rate in biological diagnostic or screeninganalysis could be increased dramatically with implementation ofthis parallel confocal excitation and detection technique. Yetanother task of this thesis was to investigations single-photongeneration by InAs-semiconductor quantum dots. We show that aquantum dots can be used for single-photon generation ondemand. Besides the single-photon generation in quantum dots,the possibility of two-photon generation, and generation ofentangled photon-pair, has also been investigated.
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| 36. |
- Blom, Hans, et al.
(författare)
-
EAST-ADL : An Architecture Description Language for Automotive Software-Intensive Systems
- 2013
-
Ingår i: Embedded Computing Systems. - Hershey : Information Science Reference. - 9781466639225
-
Bokkapitel (refereegranskat)abstract
- EAST-ADL is an Architecture Description Language (ADL) initially defined in several European-funded research projects and subsequently refined and aligned with the more recent AUTOSAR automotive standard. It provides a comprehensive approach for defining automotive electronic systems through an information model that captures engineering information in a standardized form. Aspects covered include vehicle features, requirements, analysis functions, software and hardware components, and communication. The representation of the system’s implementation is not defined in EAST-ADL itself but by AUTOSAR. However, traceability is supported from EAST-ADL’s lower abstraction levels to the implementation level elements in AUTOSAR. In this chapter, the authors describe EAST-ADL in detail, show how it relates to AUTOSAR as well as other significant automotive standards, and present current research work on using EAST-ADL in the context of fully-electric vehicles, the functional safety standard ISO 26262, and for multi-objective optimization.
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| 37. |
- Blom, Hans, et al.
(författare)
-
Electrostatic Interactions of Fluorescent Molecules with Dielectric Interfaces Studied by Total Internal Reflection Fluorescence Correlation Spectroscopy
- 2010
-
Ingår i: International Journal of Molecular Sciences. - : MDPI. - 1661-6596 .- 1422-0067. ; 11:2, s. 368-406
-
Tidskriftsartikel (refereegranskat)abstract
- Electrostatic interactions between dielectric surfaces and different fluorophoresused in ultrasensitive fluorescence microscopy are investigated using objective-based TotalInternal Reflection Fluorescence Correlation Spectroscopy (TIR-FCS). The interfacialdynamics of cationic rhodamine 123 and rhodamine 6G, anionic/dianionic fluorescein,zwitterionic rhodamine 110 and neutral ATTO 488 are monitored at various ionic strengthsat physiological pH. As analyzed by means of the amplitude and time-evolution of theautocorrelation function, the fluorescent molecules experience electrostatic attraction orrepulsion at the glass surface depending on their charges. Influences of the electrostaticinteractions are also monitored through the triplet-state population and triplet relaxationtime, including the amount of detected fluorescence or the count-rate-per-moleculeparameter. These TIR-FCS results provide an increased understanding of how fluorophoresare influenced by the microenvironment of a glass surface, and show a promising approachfor characterizing electrostatic interactions at interfaces.
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| 38. |
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| 39. |
- Blom, Hans, et al.
(författare)
-
Flocculate removal after alkaline lysis in plasmid DNA production.
- 2010
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Ingår i: Vaccine. - : Elsevier BV. - 0264-410X .- 1873-2518. ; 29:1, s. 6-10
-
Tidskriftsartikel (refereegranskat)abstract
- Alkaline lysis is the most commonly used method following harvest of bacterial cells for production of plasmid DNA. The method was originally developed for laboratory scale experiments and has shown to be challenging at larger scales. A major problem prior to further downstream processing is the risk of filter clogging without efficient removal of the flocculate that occurs after neutralization. For this purpose we here present a scalable method where the clarification of alkaline lysate is greatly simplified. Through a rapid procedure, involving the addition of ammonium hydrogen carbonate to the neutralized alkaline lysate, the flocculate is lifted to the surface of the solution by the released carbon dioxide and ammonium. After this step a clarified solution can be drained from the bottom of the vessel. The procedure does not impact pH, plasmid DNA concentration or the ratio of open circular to supercoiled plasmid DNA in the solution.
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| 40. |
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| 41. |
- Blom, Hans, et al.
(författare)
-
Fluorescence fluctuation spectroscopy in reduced detection volumes
- 2006
-
Ingår i: Current Pharmaceutical Biotechnology. - : Bentham Science Publishers Ltd.. - 1389-2010 .- 1873-4316. ; 7:1, s. 51-66
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Forskningsöversikt (refereegranskat)abstract
- Fluorescence fluctuation spectroscopy is a versatile technique applied to in vitro and in vivo investigations of biochemical processes Such as interactions, mobilities or densities with high specifity and sensitivity. The prerequisite of this dynamical fluorescence technique is to have, at a time, only few fluorescent molecules in the detection volume in order to generate significant fluorescence fluctuations. For Usual confocal fluorescence microscopy this amounts to a useful concentration in the nanomolar range. The concentration of many biomolecules in living cell or on cell membranes is, however, often quite high, usually in the micro- to the millimolar range. To allow fluctuation spectroscopy and track intracellular interaction or localization of single fluorescently labeled biomolecules ill Such crowded environments, development of detection volumes with nanoscale resolution is necessary. As diffraction prevents this in the case of light microscopy, new (non-invasive) optical concepts have been developed. In this mini-review article we present recent advancements, implemented to decrease the detection volume below that of normal fluorescence microscopy. Especially, their combination with fluorescence fluctuation spectroscopy is emphasized.
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| 42. |
- Blom, Hans, et al.
(författare)
-
Lorentzian spatial intensity distribution in one-photon fluorescence correlation spectroscopy
- 2009
-
Ingår i: Applied Optics. - 1559-128X .- 2155-3165. ; 48:31, s. 6050-6058
-
Tidskriftsartikel (refereegranskat)abstract
- The theory of autocorrelation-function evaluation in fluorescence correlation spectroscopy is applied to a Lorentzian intensity distribution. An analytical solution to the autocorrelation function for diffusion is deduced for this spatial distribution. Experimental investigation of the distribution is performed using an enlarged detector aperture in a standard confocal setup. The data from the experiment are fitted to the derived autocorrelation function, and a reasonable estimate of the spatial distribution is provided. Estimates are also compared to values computed by molecular detection efficiency simulation. The use of Lorentzian intensity distributions complements conditions where a Gaussian intensity distribution applies, expanding the applicability range of analytical correlation functions.
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| 43. |
- Blom, Hans, et al.
(författare)
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Nanoscopy-imaging life at the nanoscale : a Nobel Prize achievement with a bright future
- 2015
-
Ingår i: Physica Scripta. - : IOP Publishing. - 0031-8949 .- 1402-4896. ; 90:10
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Tidskriftsartikel (refereegranskat)abstract
- A grand scientific prize was awarded last year to three pioneering scientists, for their discovery and development of molecular 'ON-OFF' switching which, when combined with optical imaging, can be used to see the previously invisible with light microscopy. The Royal Swedish Academy of Science announced on October 8th their decision and explained that this achievement-rooted in physics and applied in biology and medicine-was awarded with the Nobel Prize in Chemistry for controlling fluorescent molecules to create images of specimens smaller than anything previously observed with light. The story of how this noble switch in optical microscopy was achieved and how it was engineered to visualize life at the nanoscale is highlighted in this invited comment.
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- Blom, Hans, et al.
(författare)
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Nearest neighbor analysis of dopamine D1 receptors and Na plus -K plus -ATPases in dendritic spines dissected by STED microscopy
- 2012
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Ingår i: Microscopy research and technique (Print). - : Wiley. - 1059-910X .- 1097-0029. ; 75:2, s. 220-228
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Tidskriftsartikel (refereegranskat)abstract
- Protein localization in dendritic spines is the focus of intense investigations within neuroscience. Applications of super-resolution microscopy to dissect nanoscale protein distributions, as shown in this work with dual-color STED, generate spatial correlation coefficients having quite small values. This means that colocalization analysis to some extent looses part of its correlative impact. In this study we thus introduced nearest neighbor analysis to quantify the spatial relations between two important proteins in neurons, the dopamine D1 receptor and Na+,K+-ATPase. The analysis gave new information on how dense the D1 receptor and Na+,K+-ATPase constituting nanoclusters are located both with respect to the homogenous (self to same) and the heterogeneous (same to other) topology. The STED dissected nanoscale topologies provide evidence for both a joint as well as a separated confinement of the D1 receptor and the Na+,K+-ATPase in the postsynaptic areas of dendritic spines. This confined topology may have implications for generation of local sodium gradients and for structural and functional interactions modulating slow synaptic transmission processes. Microsc. Res. Tech., 2011.
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