Our facilities include optical superresolution microscopes using variants of single molecule localisation microscopy such as dSTORM, structured illumination microscopy (SIM) 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:
Microscopy | Sensing | Molecular and Cell Biological Facilities
Microscopy
Confocal imaging station for multi-parameter imaging (TCSPC)
We have developed a confocal microscopy based platform integrated with a time correlated single photon counting (TCSPC) module, for fluorescence lifetime imaging measurements. The system employs either a pulsed femtosecond laser (for two-photon (2P) measurements) or a supercontinuum laser, for excitation at repetition rates ranging 2-80 MHz. Hence, it is unique in terms of its flexibility of use and sensitivity, especially for biological imaging. For more information, please contact Sofia Kapsiani (sk2067@cam.ac.uk). :
- 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 time-gated fluorescence lifetime imaging microscope (TG-FLIM)
We have developed a widefield time-gated FLIM microscope capable of taking FLIM images rapidly, as fast as 2Hz. The system is equipped with an automated stage and our home-built LabView script allows automation and unsupervised, rapid screening of samples. For more information, please contact Edward Ward (ew535@cam.ac.uk). More details on our system can also be found in:
- Laine et al., “Fast Fluorescence Lifetime Imaging Reveals the Aggregation Processes of α-Synuclein and Polyglutamine in Aging Caenorhabditis elegans.” (2019)
Selective Plane Illumination Microscope (SPIM)
We have developed a state-of-the-art SPIM microscope for high speed single-cell imaging. The system is capable of recording up to 60 fluorescence sections per second and is mostly used for live single-cell and expanded sample imaging. For more information please contact Dr Liuba Dvinskikh (lad62@cam.ac.uk).
Direct Stochastic Optical Reconstruction Microscopy (dSTORM)
We have a custom built dSTORM microscope, capable of simultaneous 2 colour superresolution imaging and we achieve an image resolution of 15 nm routinely. For more information please contact Marius Brockhoff (mb2315@cam.ac.uk). 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)
DNA Points Accumulation for Imaging in Nanoscale Topography (DNA-PAINT)
We are using a custom built DNA-Paint setup, capable of multiplexed superresolution imaging with a resolution of about 40 nm. We are currently working on several approaches to enhance acquisition speed and multi-channel imaging. For more information please contact Francesca van Tartwijk (fwv21@cam.ac.uk).
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 operate in TIRF or widefield SIM modes with simultaneous multicolour excitation - allowing the system to be flexibly adapted to the imaging problem. For details on this system please contact Dr Edward Ward (ew535@cam.ac.uk), view our super-resolution article or view our relevent publications:
- Young LJ, Ströhl F & Kaminski CF "A guide to structured illumination TIRF microscopy at high speed with multiple colors"(2016)
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 increase 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 Edward Ward (ew535@cam.ac.uk) or view our super-resolution article.
Atomic Force Microscopy
The group also has a state-of-the-art atomic force microscope (AFM) for life science and materials science research. AFM exploits the use of sharp probes for the creation of ultra-high resolution maps of surface topography alongside physical properties maps of the sample, such as elasticity, adhesion etc. The instrument in our lab, (a Bruker Resolve AFM) can be easily integrated with most inverted microscope frames. This allows for the development of correlative microscopy platforms, such as AFM-STED, AFM-FLIM and AFM-SIM. For more information please contact Dr Ioanna Mela (im337@cam.ac.uk) or view our relevent publications:
- Poudel C et al., "Correlative AFM-FLIM Measurements in Living Cells, Tissues and in Solar Cell materials" (2019)
- Curry, N. et al., "Correlative STED and Atomic Force Microscopy on Live Astrocytes Reveals Plasticity of Cytoskeletal Structure and Membrane Physical Properties during Polarized Migration" (2017)
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.