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

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Sampling frequency in livecell image acquisitionThe optimal speed of acquisition is determined by thebiological process in questionExampleCa2+ sparksVesicle movementMitosisApoptosisWoundhealingTimescaleMillisecondsSecondsMinutesHoursTens of hoursHigh sampling frequency (timedomain) requires simplification of the experiment and theacquisition protocol– Preferably very few mechanical parts involved– Fast shutters for illumination– Sensitive detectors for acquisitionFluorescence imaging 1
Confocal vs. widefield microscopesSelection between confocal and widefield microscopesConfocal if high resolution spatialinformation is required More flexibility in bleachingapplicationsWidefield if high resolution temporalinformation required More sensitive in lowlight detectionIn both cases acquisition is usuallybased on monochromaticdetectors –PMTs or CCD camerasMulticolour detection: Confocals: selection of excitation wavelength restricted ie. laser lines Widefield systems: selection of emission wavelength more restricted ie. filters Both are equally appropriate for multichannel/ multicolor detectionFluorescence 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 and 8: Phototoxicity and photobleaching of
Page 9 and 10: Fluorochromes and probes for live c
Page 11: Objective lenses -resolution and se
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

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