Fluorescence lifetime imaging microscopy (FLIM)

The Wolfson Bioimaging Facility provides fluorescence lifetime imaging microscopy by addition of specialised equipment and software to one of its Leica SP8 CLSM systems.

Monitoring the fluorescence lifetime of a fluorophore (the average time between excitation and photon emission) rather than its intensity provides concentration-independent information on local microenvironment.

Potential FLIM applications include FRET (Fluorescence Resonant Energy Transfer) analysis of fluorophore interactions, measurement of ions and signalling molecules, and analysis of the physical environment of the fluorophore.

Our FLIM system is part of the Leica SP8X system also equipped for STED and FCS and uses a pulsed white light laser which can be tuned precisely within 470-670nm excitation range. Specialised single molecule detectors and SymPhoTime software (PicoQuant) facilitate TSCPC (time correlated single photon counting) FLIM.

Although FLIM acquisition is relatively straightforward with our system, FLIM experiments require specialised expertise and are relatively analysis-intensive.

STED/FLIM/FCS technical specifications (PDF, 179kB)

Recent publications including data acquired with our FLIM system:

Tiwari, AlibhaiFermin (2018) Above 600 mV Open-Circuit Voltage BiI3 Solar Cells. ACS Energy Lett doi 10.1021/acsenergylett.8b01182

Ross, Bridges, Fletcher, Shoemark, Alibhai, Bray, Beesley, Dawson, Hodgson, Mantell, Verkade, Edge, Sessions, Tew & Wolfson (2017) Decorating self-assembled peptide cages with proteins. ACS Nano Jul 11 doi: 10.1021/acsnano.7b02368

Baarlink, Plessner, Sherrard, Morita, Misu, Virant, Kleinschnitz, Hatniman, Alibhai, Baumeister, Miyamaoto, Endesfelder, Kaidi & Grosse (2017) A transient pool of nuclear F-actin at mitotic exit controls chromatin organization. Nature Cell Biology doi: 10.1038/ncb3641

More information and access

For further information or to arrange access to this equipment, contact one of the team.

We welcome comments or suggestions. Please contact one of the team or one of the steering group.

Mouse cell expressing fluorescently tagged histone H2B (green and red) and analysed using FLIM-FRET to determine the degree of genome/chromatin compaction, as depicted. (Courtesy of Abderrahmane Kaidi’s Nuclear Dynamics Laboratory)
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