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Authors = B. Christoffer Lagerholm

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6 pages, 880 KiB  
Brief Report
Using kICS to Reveal Changed Membrane Diffusion of AQP-9 Treated with Drugs
by Jakob L. Kure, Thommie Karlsson, Camilla B. Andersen, B. Christoffer Lagerholm, Vesa Loitto, Karl-Eric Magnusson and Eva C. Arnspang
Membranes 2021, 11(8), 568; https://doi.org/10.3390/membranes11080568 - 28 Jul 2021
Cited by 3 | Viewed by 2290
Abstract
The formation of nanodomains in the plasma membrane are thought to be part of membrane proteins regulation and signaling. Plasma membrane proteins are often investigated by analyzing the lateral mobility. k-space ICS (kICS) is a powerful image correlation spectroscopy (ICS) technique and a [...] Read more.
The formation of nanodomains in the plasma membrane are thought to be part of membrane proteins regulation and signaling. Plasma membrane proteins are often investigated by analyzing the lateral mobility. k-space ICS (kICS) is a powerful image correlation spectroscopy (ICS) technique and a valuable supplement to fluorescence correlation spectroscopy (FCS). Here, we study the diffusion of aquaporin-9 (AQP9) in the plasma membrane, and the effect of different membrane and cytoskeleton affecting drugs, and therefore nanodomain perturbing, using kICS. We measured the diffusion coefficient of AQP9 after addition of these drugs using live cell Total Internal Reflection Fluorescence imaging on HEK-293 cells. The actin polymerization inhibitors Cytochalasin D and Latrunculin A do not affect the diffusion coefficient of AQP9. Methyl-β-Cyclodextrin decreases GFP-AQP9 diffusion coefficient in the plasma membrane. Human epidermal growth factor led to an increase in the diffusion coefficient of AQP9. These findings led to the conclusion that kICS can be used to measure diffusion AQP9, and suggests that the AQP9 is not part of nanodomains. Full article
(This article belongs to the Special Issue Dynamics and Nano-Organization in Plasma Membranes)
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7 pages, 984 KiB  
Article
Defining the Diffusion in Model Membranes Using Line Fluorescence Recovery after Photobleaching
by Jakob L. Kure, Camilla B. Andersen, Thomas E. Rasmussen, B. Christoffer Lagerholm and Eva C. Arnspang
Membranes 2020, 10(12), 434; https://doi.org/10.3390/membranes10120434 - 17 Dec 2020
Cited by 9 | Viewed by 3552
Abstract
In this study, we explore the use of line FRAP to detect diffusion in synthetic lipid membranes. The study of the dynamics of these membrane lipids can, however, be challenging. The diffusion in two different synthetic membranes consisting of the lipid mixtures 1:1 [...] Read more.
In this study, we explore the use of line FRAP to detect diffusion in synthetic lipid membranes. The study of the dynamics of these membrane lipids can, however, be challenging. The diffusion in two different synthetic membranes consisting of the lipid mixtures 1:1 DOPC:DPPC and 2:2:1 DOPC:DPPC:Cholesterol was studied with line FRAP. A correlation between diffusion coefficient and temperature was found to be dependent on the morphology of the membrane. We suggest line FRAP as a promising accessible and simple technique to study diffusion in plasma membranes. Full article
(This article belongs to the Special Issue Dynamics and Nano-Organization in Plasma Membranes)
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19 pages, 3570 KiB  
Article
Exploring the Potential of Airyscan Microscopy for Live Cell Imaging
by Kseniya Korobchevskaya, B. Christoffer Lagerholm, Huw Colin-York and Marco Fritzsche
Photonics 2017, 4(3), 41; https://doi.org/10.3390/photonics4030041 - 7 Jul 2017
Cited by 67 | Viewed by 29987
Abstract
Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment [...] Read more.
Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment principle in order to enhance both the spatial resolution and signal-to-noise-ratio without increasing the excitation power and acquisition time. Here, we present a detailed study evaluating the performance of Airyscan as compared to confocal microscopy by imaging a variety of reference samples and biological specimens with different acquisition and processing parameters. We found that the processed Airyscan images at default deconvolution settings have a spatial resolution similar to that of conventional confocal imaging with a pinhole setting of 0.2 Airy Units, but with a significantly improved signal-to-noise-ratio. Further gains in the spatial resolution could be achieved by the use of enhanced deconvolution filter settings, but at a steady loss in the signal-to-noise ratio, which at more extreme settings resulted in significant data loss and image distortion. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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