Although their composition varies, one of the lipids often associated with lipid nano-domains is Phosphatidyl Inositol 4,5 Bisphosphate (PIP 2). Lipid rafts are defined as small (10–200 nm) heterogeneous sterol and sphingolipid enriched domains that compartmentalize cellular processes ( Pike, 2006). In turn, this allows the localization of individual active fluors to be achieved with very high precision, and repeated cycling of the fluors in a sample allows a very high resolution image to be built up over time. In an appropriate buffer, organic fluorophores can bleach reversibly (“blinking”), which allows a small, random proportion of the fluors in a field to be active at one time. PhotoActivated Light Microscopy (PALM) ( Manley et al., 2008) and the conceptually similar Direct Stochastic Optical Reconstruction Microscopy (dSTORM) ( Rust et al., 2006) are other widefield super-resolution techniques that are considerably simpler in their equipment requirements. Techniques which improve on this include STimulated Emmission Depletion (STED) microscopy which is a point scanning technique like a confocal, but shrinks the focal volume by generated a depleted region around the point of interest to sharpen the resolution ( Hein et al., 2008), and Structured Illumination Microscopy (SIM) which analyses the moire interference patterns due to application of non-uniform illumination ( Gustafsson et al., 2008). Most subcellular processes occur in regions below the diffraction limit (approximately 200 nm under ideal conditions). Natural Sciences Identifiers URN: urn:nbn:se:mau:diva-4263 DOI: 10.1016/j.colsurfb.2018.09.031 ISI: 000454377300025 Local ID: 26667 OAI: oai:DiVA.Over the last few years the development of super-resolution imaging techniques has begun a revolution in cell biology ( Huang et al., 2010). Neutron Reflectometry, Phosphoinositides, Supported lipid bilayers National Category Place, publisher, year, edition, pagesElsevier, 2019. These results support the proposed PIP3 tendency to interact with the other lipid headgroups as PC, so far exclusively suggested by MD simulations. The headgroup layer thicknesses of the pure POPC and the mixed POPC/DOPIP3 bilayer suggest that the DOPIP3-headgroups have a preferred orientation, which is not perpendicular to the membrane surface, but instead it is close to the surrounding lipid headgroups. The two bilayer leaflets showed the same DOPIP3 concentration, thus suggesting the formation of a symmetric bilayer. On the other hand, NR was used to collect experimental structural data on the DOPIP3 location and orientation within the lipid membrane. Specifically, QCM-D and AFM were used to identify the best condition for lipid deposition and to estimate the total bilayer thickness. Quartz Crystal Microbalance with Dissipation monitoring (QCM-D), Atomic Force Microscopy (AFM) and Neutron Reflectometry (NR), provided detailed information on the formation of the SLB and the location of DOPIP3 in the lipid membrane. For this purpose, supported lipid bilayers (SLBs) containing 1,2-dioleoyl-sn-glycero-3-phospho-(1'-myo-inositol-3',4',5'-trisphosphate (DOPIP3) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were used as mimics of biomembranes. The present study aims to reveal the structure of PIP-containing lipid membranes, which so far has been little explored. ![]() ![]() Among all, phosphoinositol-3',4',5'-trisphosphate (PIP3) is a secondary signaling messenger that regulates the function of proteins involved in cell growth and gene transcription. Phosphoinositide (PIP) lipids are anionic phospholipids playing a fundamental role for the activity of several transmembrane and soluble proteins. 202-209 Article in journal (Refereed) Abstract Show others and affiliations 2019 (English) In: Colloids and Surfaces B: Biointerfaces, ISSN 0927-7765, E-ISSN 1873-4367, Vol.
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