For more information please contact: Dr. Chris Tynan.
Instrument and technique summary
|Technique and applications||Specifications|
Total Internal Reflection (SM-TIRF) Microscopy Widefield multidimensional fluorescence imaging where signal is limited to 100 nm layer adjacent to coverslip.
Biological and medical research, material science, chemistry
Lasers: 405, 488, 532, 561, 642 nm or white-light excitation. Three-colour simultaneous imaging.
Detectors: Andor iXon EMCCDs.
Objectives: 100x TIRF oil immersion.
Extra information: Excitation and emission polarisation resolvable.
molecule fluorescence sensitivity is achieved by using Total Internal
Reflection Fluorescence (TIRF) or High Inclination illumination to
restrict background fluorescence, using extremely sensitive cooled
EMCCDs and judicious sample preparation to minimise non-specific probe
binding and molecular crowding in the sample.
molecule fluorescence imaging allows a target to be studied without
ensemble averaging of its properties. It can be applied to a range of
commercially available fluorescent dyes, commonly used fluorescent
proteins and quantum dots and applied to a wide range of sample types.
It is particularly useful for investigating biological systems and
OCTOPUS single molecule microscopes have been heavily utilised for
studying the behaviour of membrane proteins in cultured cells.
single molecule microscopes are flexible, multidimensional instruments.
This means that they can be adapted for use with any technique that
requires multiple image channels resolved by fluorescence wavelength or
Multi-channel TIRF image of HeLa cells
Epi-fluorescence, Total Internal Reflection Fluorescence (TIRF)
Illumination/fluorescence excitation sources
405nm, 488nm, 561nm, 640nm continuous wave excitation
Available channels and detectors
1 - 6 channels split by wavelength. Andor cooled EMCCDs. Maximum frame rate 50 Hz (faster frame rates are possible by reducing the effective chip size).
Multicolour TIRF microscopy, Single Particle Tracking (SPT), Fluorescence Localisation Imaging with Photobleaching (FLImP), Single Molecule Förster Resonance Energy Transfer (SM-FRET), combined wavelength and polarisation resolved single molecule imaging.
Single molecule tracks reveal co-localisation of fluorescently labelled features
Samples can created in glass bottom dishes or mounted on coverslips. Biological samples can be live or chemically fixed. Temperature control is available in the range 18-40°C.