Light & Laser Microscopy

INSTRUMENTS LOCATED AT CHARLES PERKINS CENTRE (CPC)

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 labels it enables individual proteins and their interactions to be localised with a lateral 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 TCS SP8 STED 3X microscope
Super Resolution Fluorescence Microscopy collects images of fluorescent molecules with resolution that is not limited by the diffraction limit of light. Stimulated Emission Depletion (STED) is built on a Confocal microscope and is a laser scanning technique.

Confocal excitation is available using a 405nm laser and Argon laser with 458, 476, 488, 496 and 514nm lines and a White Light Tunable pulsed laser 470-670nm that can be used for gated STED. In addition to the excitation laser, STED scans a second laser line at 592nm, improving lateral resolution to a theoretical limit of 50nm, actual ~100nm. This STED laser can also be manipulated to improve axial resolution to a theoretical limit of 130nm, actual ~300nm. If optimal resolution in XY and Z is required the actual achievable resolution is approximately 150nm laterally and 500nm axiallly.

This microscope also features an Insight DeepSee laser for multiphoton and Second Harmonic Generation (SHG) imaging with excitation wavelenghts of 680-1300nm. Matched PMTs for forward and backward SHG. The two external HyD detectors collect more scattered light from thick samples especially with long distance working objectives (25x W NA 0.95 objective with WD 2.0mm and 40x W NA 1.1 objective with WD 610um).

Black paneled box incubator around microscope for live cell imaging.

Specialists: Minh Huynh and Pam Young

   
     

Nikon A1 Advanced Confocal microscope
Advanced Confocal for Spectral Scanning, FRAP, FRET, FLIM, FCS, FCCS in live cells.

Specialists: Ellie Kable and Pam Young

 
     

Nikon TIRF & DMD microscope
Total Internal Reflection microscopy is a Super Resolution technique.

Specialists: Minh Huynh and Pam Young
 
     

Nikon Ti-E Spinning Disk Confocal Live Cell microscope
Live cell fluorescence, DIC, large mosaics, multi-point, time series, perfect focus.

Specialists: Minh Huynh and Pam Young
 
     

Nikon C2 Confocal microscope
Upright Confocal microscope with motorised stage.

Specialists: Minh Huynh and Pam Young
 
     

Nikon Ni_E Widefield microscope
Upright microscope for mosaics and multipoint, using colour camera for histology or mono camera for fluorescence.

Specialists: Minh Huynh and Pam Young
 
     

Leica DM6000 microscope
Upright microscope for mosaics using colour camera for histology and fluorescence.

Specialists: Minh Huynh and Pam Young
 
     

Zeiss Shuttle & Find microscope
Correlative Fluorescence with SEM on Zeiss Sigma.

Specialists: Minh Huynh and Pam Young
 
     

Zeiss Axiovert 200M microscope
Inverted Fluorescence microscope for FURA2 imaging.

Specialists: Minh Huynh and Pam Young
 
     

Olympus IX71 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.

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

INSTRUMENTS LOCATED AT MADSEN BUILDING

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 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

   FV1000
   

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
     

Leica DM6000 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
   
     

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: Takanori Sato 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