HSØ core facility for confocal microscopy
Confocal microscopy is a further development of fluorescence microscopy, a methodology known for its high sensitivity for detection of specific molecules in biological specimens. When compared with conventional fluorescence microscopy, confocal microscopy offers superior resolution and contrast. This is achieved through the use of laser-based point illumination and a spatial pinhole that eliminates out-of-focus light. Because of its excellent spatial resolution, confocal microscopy is well suited for imaging optical sections of semi-thick (up to 60 ?m) samples, and such optical sections can be digitally reconstructed into 3D projections. The point illumination process is sufficiently fast to allow imaging of live cells and tissues, and the confocal microscopes at the HSØ core facility are equipped with climate chambers that ensure long-term viability of live specimens. The combination of laser illumination with appropriate excitation and emission filters makes modern confocal microscopes ideally suited for multiple labelling of biological specimens, and quadruple labelling is achievable with all the HSØ core facility confocal microscopes. The ability to visualize up to four different molecules in the same specimen is a great advantage for many applications. While fluorescence microscopy has classically employed fluorescently labelled secondary antibodies for visualization of specific antigens, modern recombinant DNA technology has offered the possibility of using additional probes such as fluorescently labelled lipid-binding proteins (some of these have been developed by the applicants), nucleic acids, various fluorescent small molecules, fusion proteins with recombinant fluorescent proteins such as GFP and mCherry, and CLIP or SNAP tags that enable in vivo labelling of specific proteins with small fluorescent dyes. These developments have greatly expanded our possibilities to label specific molecules in cells and tissues and have facilitated the use of live imaging methods. Although confocal microscopes are still mostly used for determinations of the intracellular localizations of specific biomolecules, additional applications are becoming increasingly important. In particular, fluorescence recovery after photobleaching (FRAP) has emerged as a very important method to monitor molecular diffusions in living cells, and fluorescence resonance energy transfer (FRET) has become an important method for detecting molecular interactions in situ. Likewise, the ability to photoactivate or photoconvert specific fluorescent probes has opened new possibilities for monitoring protein dynamics in living cells. The HSØ core facility offers these methodologies, which have a broad spectrum of applications. In 2015 the core facility has served more than 200 users at HSØ and contributed to 20 Scientific publications, some of these in very prestigious journals. Overall the facility has contributed significantly to the success of HSØ and Oslo University Hospital as research institutions.
Kjernefasiliteten bidrar positivt til en rekke forskningsprosjekter innen sentrale sykdomsområder som kreft, autoimmunitet og hjerte-kar-lidelser, og den bidrar til å heve nivået på slik forskning innen HSØ. Legger man til grunn at økt kvalitet på forskning fører til økt kvalitet på pasientbehandling, skal derfor kjernefasiliteten ha positive konsekvenser for helsetjenesten innen HSØ. .
Flotillin depletion affects ErbB protein levels in different human breast cancer cells.
Biochim Biophys Acta 2014 Sep;1843(9):1987-96. Epub 2014 apr 18
The ether lipid precursor hexadecylglycerol protects against Shiga toxins.
Cell Mol Life Sci 2014 Nov;71(21):4285-300. Epub 2014 apr 18
Photochemical activation of the recombinant HER2-targeted fusion toxin MH3-B1/rGel; Impact of HER2 expression on treatment outcome.
J Control Release 2014 May 28;182():58-66. Epub 2014 mar 15
Photoactivation approaches reveal a role for Rab11 in FGFR4 recycling and signalling.
Traffic 2014 Jun;15(6):665-83. Epub 2014 mar 31
Cell density-induced changes in lipid composition and intracellular trafficking.
Cell Mol Life Sci 2014 Mar;71(6):1097-116. Epub 2013 aug 7
Vps11, a subunit of the tethering complexes HOPS and CORVET, is involved in regulation of glycolipid degradation and retrograde toxin transport.
Commun Integr Biol 2014 Jan 1;7(1):e28129. Epub 2014 apr 3
The novel EpCAM-targeting monoclonal antibody 3-17I linked to saporin is highly cytotoxic after photochemical internalization in breast, pancreas and colon cancer cell lines.
MAbs 2014 Jul-Aug;6(4):1038-50.
PIKfyve, MTMR3 and their product PtdIns5P regulate cancer cell migration and invasion through activation of Rac1.
Biochem J 2014 Aug 1;461(3):383-90.
Association of CHMP4B and autophagy with micronuclei: implications for cataract formation.
Biomed Res Int 2014;2014():974393. Epub 2014 mar 11
CK2 involvement in ESCRT-III complex phosphorylation.
Arch Biochem Biophys 2014 Mar 1;545():83-91. Epub 2014 jan 15
ANCHR mediates Aurora-B-dependent abscission checkpoint control through retention of VPS4.
Nat Cell Biol 2014 Jun;16(6):550-60. Epub 2014 mai 11
Monitoring phosphatidylinositol 3-phosphate in multivesicular endosome biogenesis.
Methods Enzymol 2014;534():3-23.
A thymic stromal lymphopoietin-responsive dendritic cell subset mediates allergic responses in the upper airway mucosa.
J Allergy Clin Immunol 2014 Sep;134(3):613-621.e7. Epub 2014 jun 21
The effect of hepatic progenitor cells on experimental hepatocellular carcinoma in the regenerating liver.
Scand J Gastroenterol 2014 Jan;49(1):99-108. Epub 2013 nov 5
Indirect CD4+ T-cell-mediated elimination of MHC II(NEG) tumor cells is spatially restricted and fails to prevent escape of antigen-negative cells.
Eur J Immunol 2014 Sep;44(9):2625-37. Epub 2014 jun 20
Computational modeling of Takotsubo cardiomyopathy: effect of spatially varying ß-adrenergic stimulation in the rat left ventricle.
Am J Physiol Heart Circ Physiol 2014 Nov 15;307(10):H1487-96. Epub 2014 sep 19
Calcium signalling in developing cardiomyocytes: implications for model systems and disease.
J Physiol 2015 Mar 1;593(5):1047-63. Epub 2015 feb 9
Hypokalaemia induces Ca2+ overload and Ca2+ waves in ventricular myocytes by reducing Na+,K+-ATPase a2 activity.
J Physiol 2014 Oct 17. Epub 2014 okt 17
Variable t-tubule organization and Ca2+ homeostasis across the atria.
Am J Physiol Heart Circ Physiol 2014 Aug 15;307(4):H609-20. Epub 2014 jun 20
A dominant STIM1 mutation causes Stormorken syndrome.
Hum Mutat 2014 May;35(5):556-64. Epub 2014 apr 9