Light & Laser Microscopy

Leica Ground State Depletion (GSD) microscope

This new Leica instrument enables researchers to see the detail of cellular systems at a level previously only achievable with electron microscopy. By using fluorescent lables it will enable individual proteins and their interactions to be localised with a resolution of 20nm.

The system is based on a fully automated Total Internal Reflection Fluorescence (TIRF) microscope with laser lines of 488, 532 and 642nm. It also has a 405nm laser for backpumping to optimise GSD data collection.

This system has recently been updated to collect 3D GSD images, overcoming the axial resolution limit, achieving down to a resolution of 50nm in the axial direction. This upgrade includes a 160x high-performance objective specifically developed for super-resolution microscopy.

Specialists: Pam Young and Minh Huynh

 

 

Leica SP5 II confocal and multiphoton microscope
This instrument covers a broad range of requirements in confocal and multiphoton imaging - with the full array of scan speeds at highest resolution. You can now image your live cells then image the same “dynamic” event later at high resolution in the TEM. The microscope is equipped with a new Spectra-Physics Mai Tai DeepSee™ Ti:Sapphire femtosecond pulsed laser, specialised objectives and external non-descanned detectors. This means that we can image more than 300 microns deep into thick specimens without any signal drop-off. Single photon excitation at 458, 476, 488, 496, 514, 561 and 633nm. Multiphoton excitation variable 690-1060nm.

The system is equipped with a resonance and galvanometer scanner, so we can now image around three times faster at higher resolution than ever before (e.g. a 512 x 128 pixel array every 15ms).

The system features Fluorescence Lifetime Imaging (FLIM) and forward and backward Second Harmonic Generation (SHG) capabilities.  It is equipped with a time-correlated single-photon-counting (TCSPC) board for (FLIM). This means that we can measure the lifetime of a fluorophore faster and more accurately than before at various excitation wavelengths. Its photomultiplier tubes (PMTs) in the forward direction are calibrated to optimise forward and backward SHG imaging.

Specialists: Ellie Kable and Pam Young
  Leica TCS SPII Multi-Photon Microscope
   

Olympus FluoView FV1000 Confocal Microscope
Equipped with 405, 458, 488, 514, 559 and 633 nm lasers and a transmitted light detector allowing for the majority of fluorophores to be imaged with a beautiful DIC (or brightfield) overlay. Fantastic spectral resolution that enables users to collect the emission profile of a visible fluorophore at any point in their sample. This gives rise to a spectral un-mixing function, which allows separation of spectrally close fluorophores, particularly handy for users with autofluorescence interfering with the signal of their labelled target of interest. The instrument allows to bleach/activate and image at the same time, due to the second scanner that is integrated in the system. Users performing FRET, FRAP, FLIP, photoactivation or uncaging studies, will find this is particularly handy. Offers scanning speed of up to 1 frame every 0.065 s, in bidirectional mode. This, combined with the heated-stage insert, results in excellent temporal resolution for the short-term imaging of living cells. 

Specialists:Pam Young and Minh Huynh

  Olympus FluoView FV1000 Confocal Microscope
   

Olympus CellR Live-Cell Microscope
Equipped with a stage-top incubator and CO2 controller, to allow the long-term imaging of cells. Ability to image extremely quickly. ZDC or 'zero drift control' ensures images stay in focus throughout the acquisition time. The microscope is also equipped with a Marzhauzer motorised stage, that with precision in X, Y and Z, and can return to a single point when imaging multiple points in a sample. Extremely stable for 24-48 hour imaging sessions.

Specialists: Pam Young and Minh Huynh

  Olympus CellR Live-Cell Microscope
   

Olympus TIRF (Total Internal Reflection Fluorescence) Microscope
TIRF is a technique employed to look at events that occur at interfaces of objects with different refractive indices. See events that occur at the cell membrane versus those that occur within the cytoplasm, something that is unachievable by traditional light microscopy techniques due to axial resolution limitations. The system is equipped with a 488 nm laser for TIRF and an anti-vibration table, along with the addition of standard filter sets, making it the most sensitive of all our fluorescent microscopes.

Specialists: Ellie Kable and Pam Young
  Olympus CellR Live-Cell Microscope
   

Nikon C1 LIMO Confocal Microscope with Fluorescence Lifetime Imaging
Confocal microscope with three detectors and 3 solid-state lasers (405nm, 488nm, 561nm) for standard confocal imaging, and picosecond pulsed 440nm laser and high-speed gated detection system for lifetime imaging. Standard filter sets plus CFP/YFP and red/far red. Inverted microscope, with standard objectives and water immersion objective optimised for violet excitation. Software for lifetime measurement, ratiometric imaging and 3D reconstruction. Used in life sciences; lifetime imaging permits Förster Resonant Energy Transfer (FRET) studies of molecular interactions. As a standard confocal the 405nm laser permits imaging of DAPI and similar nuclear stains.

Example image:
Upper part - 3-channel confocal image of coral Acropora millepora showing fluorescent pigments (green) and symbiotic algae (red). Lower picture fluorescence lifetime images of the coral pigments in the same field of view.

Specialists: Ellie Kable and Pam Young

  Nikon C1 LIMO Confocal Microscope with Fluorescence Lifetime Imaging
   

Nikon Eclipse E800 Fluorescence Microscope
General-use light and fluorescence microscope for life sciences, material science and medical applications. Standard filter sets (FITC, GFP, Rhodamine, DAPI) for fluorescence. Bright field, phase and Nomarski optics.

Leica DFC400 camera with 1.4 megapixel for very fast image captures.

Example image:
Prepared fluid mount of Spirogyra showing Nomarski optics and fluorescence using three standard filter sets (FITC, Rhodamine and DAPI).

Specialists: Ellie Kable and Pam Young

  Nikon Eclipse E800 Fluorescence Microscope (SensiCam)
   
   

Olympus BX61 Microscope (SIS)

Reflected and transmitted light microscope. Image analysis system for material and life sciences. Useful for thick specimens. Also operates as a fluorescence microscope.

Specialists: Adam Sikorski and Pam Young
  Olympus BX61 Motorized System Microscope (SIS)

Olympus BX60 Light Microscope

General use light microscope for life sciences, materials science and medical applications. Bright field and reflected light microscopy.

Specialist: Pam Young