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SUMMARY:Applications of Fluorescence Lifetime Imaging Microscopy (FLIM) fo
 r Alzheimer's Disease and Stem Cell research - Dr Colin Hockings\, Departm
 ent of Chemical Engineering and Biotechnology
DTSTART:20190213T130000Z
DTEND:20190213T140000Z
UID:TALK114487@talks.cam.ac.uk
CONTACT:Dr Chris Ness
DESCRIPTION:The fluorescence lifetime of a fluorophore is affected by its 
 local environment\, but is relatively independent of its concentration. Ch
 anges in viscosity\, temperature\, ion or oxygen concentration\, as well a
 s the presence of other fluorophores (FRET) can be accurately measured. In
  this seminar\, I will show how we have developed FLIM assays in live cell
 s to measure protein aggregation\, nuclear compaction\, and to improve the
  detection of low protein concentrations. These assays have been crucial t
 o two projects:\n\nIn Alzheimer's Disease\, Tau aggregates spread through 
 the brain\, but it is not yet clear what triggers Tau aggregation nor how 
 these aggregates are transferred between neurons. We have found that extra
 cellular monomeric Tau can be taken up by neurons\, and the low pH of the 
 lysosomal compartment is necessary and sufficient to trigger its aggregati
 on. This aggregated Tau in endosomes and lysosomes can be transferred betw
 een neurons via two different mechanisms – via axonal connections\, or s
 ecreted into the media. We propose that aggregated Tau in lysosomes consti
 tutes a decision point between its degradation\, its secretion into the ex
 tracellular space\, or its anterograde transmission via synapses. Treatmen
 ts that affect lysosome physiology may increase the safe disposal of aggre
 gated Tau and reduce its prion-like transmission.\n\nCurrent methods to st
 udy genome compaction in live cells by fluorescence microscopy require the
  expression of fluorescent proteins. Using a new fluorescence lifetime ass
 ay\, we could confirm increased nuclear compaction in Nanog-/- embryonic s
 tem (ES) cells. We have found that naïve ES cells have a partially compac
 ted genome\, supporting the hypothesis that ES cells must transit through 
 a 'primed' de-compacted state before committing to differentiation.
LOCATION:Dept Chemical Engineering and Biotechnology\, Philippa Fawcett Dr
 ive CB3 OAS (Lecture Theatre 3\, Level 3 )
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