Our facilities include optical superresolution microscopes using variants of single molecule localisation microscopy such as dSTORM, structured illumination microscopy systems and stimulated emission depletion (STED) microscope systems. Furthermore we have systems for whole organism imaging methods such as selective plane illumination microscopy. These efforts were funded over the past 5 years by grants from the Wellcome Trust, MRC, EPSRC, BBSRC and the Alzheimer Research Trust UK. We have state of the art laser sources and detector systems including several supercontinuum laser sources, Ti:Sa lasers, EMCCD and scientific grade CMOS cameras, microscopic imaging stations etc. We also have state of the art molecular biology, cell culture and chemical preparation laboratories. Key equipment is listed below:
Confocal imaging station for multi-parameter imaging
We have developed over the last 10 years a confocal microscopy platform capable of measuring excitation spectrum, emission spectrum, fluorescence polarisation and Fluorescence lifetime using time correlated single photon counting (TCSPC). The system employs a pulsed supercontinuum laser source for excitation and is unique in terms of flexibility, sensitivity, and flexibility of operation. The system is described in more detail here and in several publications:
- Frank JH et al., "A white light confocal microscope for spectrally resolved multidimensional imaging" (2007)
- Kaminski CF et al., "Supercontinuum radiation for applications in chemical sensing and microscopy" (2008)
- Esposito A et al., "Design and application of a confocal microscope for spectrally resolved anisotropy imaging" (2011)
Widefield frequency domain fluorescence lifetime imaging microscope (Lambert Instruments LIFA and LaVision Picostar systems)
More details on our system are found here and in the following publications:
- Elder AD et al., "Calibration of a wide-field frequency-domain fluorescence lifetime microscopy system using light emitting diodes as light sources" (2006)
Selective Plane Illumination Microscope (SPIM)
We have developed a state-of-the-art SPIM microscope capable of one and two photon excitation, confocal slit scanning to reduce out of focus image contamination, adaptive optics to shape the illumination beam for deep tissue penetration, and a capability to perform one and two photon excitation. The system is capable of recording up to 60 fluorescence sections per second and is mostly used for deep tissue live embryo imaging (drosophila, zebrafish, C. elegans and organotypic slices).
Direct Stochastic Optical Reconstruction Microscopy (dSTORM)
We have a custom built dSTORM microscope, capable of simultaneous 2 colour superresolution imaging and we achieve an imageresolution of 15 nm routinely. For details on this system and representative work see:
- Kaminski Schierle GS et al., "In Situ Measurements of the Formation and Morphology of Intracellular ß-Amyloid Fibrils by Super-Resolution Fluorescence Imaging" (2011)
- Rees EJ et al., "Blind Assessment of Localisation Microscope Image Resolution" (2012)
- Erdelyi M et al., "Correcting chromatic offset in multicolor super-resolution localization microscopy" (2013)
Structured Illumination Microscope (SIM)
In our lab we have developed a SIM microscope capturing super-resolution images with, a resolution twice as high as for confocal imaging, in fractions of a second. The system can be flexibly adapted to a range of SIM modes and uses a supercontinuum laser for multicolour excitation - allowing the system to be flexibly adapted to the imaging problem. For details on this system please contact Laurie Young (firstname.lastname@example.org) or view our super-resolution article.
Stimulated Emission Depletion Microscope (STED)
We have developed a state of the art STED microscope for fast imaging of samples labelled with red dyes such as the ATTO or Aberrior dyes. The STED system uses a Ti:Sa laser beam which is spatially shaped by an SLM to reduce the effective resolution of a standard confocal microscope from 250 nm to 30-90 nm. This technique works at reasonable imaging speed and depths and works well to image molecules deep within cells. For more information please contact Pierre Mahou (email@example.com) or view our super-resolution article.
Leica SP5 confocal imaging station with resonant scanner
This is a variant of optical tweezers using counterpropagating laser beams across microfluidic channels. This permits trapping of biological objects (e.g. cells) and measurement of their viscoelastic properties. For representative work see:
- Mauritz J et al., "Detection of Plasmodium falciparum infected red blod cells by optical stretching" (2010)
- Liquid phase cavity enhanced absorption spectrometer (CEAS)
- Ultra high sensitivity gas phase cavity ring down (CRDS) and CEAS spectrometer employing supercontinuum radiation.
- Several supercontinuum and fibre laser sources, Nd:YAG lasers, microchip lasers, intensified and non-intensified CCD cameras, avalanche photodiodes, spectrometers, etc.
Molecular and cell biological facilities
We have state of the art molecular biology, class II cell culture and nematode facilities. The facilities include equipment to perform protein purification, molecular cloning, bio-chemical and -physical assays, and mammalian cell transfection.