Live cell imaging - Biomedicum Imaging Unit (BIU) - Helsinki.fi

More documents

Recommendations

Info

Fluorescence generationJablonski diagram –exploitation of physicsInternal conversion andvibrational relaxation1psSinglet stateBleachingdecompositionIntersystem crossingFRAPanalysesS1TripletTwophotonexcitationFluorescenceemission 1 nsTriplet stateEnergyExcitation/absorption 1fsPhosphorencedelayed > ushrsFRETemission
Fluorochromes and probes for live cell imagingAntibodies, fluorescent probes, GFP variantsConjugated antibodies –highly specific• Used for extracellular domains ie. receptors• May allow quantitation of single receptorsSynthetic fluorescently tagged compounds• Designed for numerous organelles and molecules• May need a strategy for membrane permeabilityGFP variants –a palette of colours• Gene expression and protein translation• Allow dynamic applications FRAP, FRET etc• No major adverse functions in mammalian cellsFluorescence imaging 1
Page 1 and 2: Seminar Series on Advanced Imaging
Page 3 and 4: Properties of fluorescence emission
Page 5 and 6: Selection of fluorochromesDecay of
Page 7: Phototoxicity and photobleaching of
Page 11 and 12: Objective lenses -resolution and se
Page 13 and 14: Confocal vs. widefield microscopesS
Page 15 and 16: Livecell applications -intensity
Page 17 and 18: 3D colocalization analysis of proco
Page 19 and 20: Volume calculations and in situ zym
Page 21 and 22: Fluorescence recovery after photobl
Page 23 and 24: Image deconvolution and restoration
Page 25: Acknowledgements and contact detail

Live cell imaging - Biomedicum Imaging Unit (BIU) - Helsinki.fi

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?