Light & Laser Microscopy

LOCATED AT CHARLES PERKINS CENTRE (CPC)


PerkinElmer Mantra microscope
Visualise, analyse, quantify and phenotype immune cells in situ.

The Mantra quantitiative pathology workstation, with inForm image analysis software, enables easy visualisation, quantification and phenotyping of cells in situ in tissue sections using multiplexed biomarkers. This integrated workstation features a spectral detector to collect spectral data for unmixing overlapping signals and removal of autofluorescence. inForm software features user-trained algorithm to enable automatic identification of specific tissue types based on tissue morphology and indentification of cells within the tissue for quantification.

Specialists: Pam Young and Ying ying Su

 

   

Leica Ground State Depletion (GSD) microscope
This Leica SR GSD 3D is a widefield fluorescence and TIRF microscope which uses ground state depletion and single molecule return to localise molecules of interest. Using this technique coupled with optimal sample preparation and supported dyes, strucutures of interest can be resolved up to 20nm laterally and 50nm axially.

Lasers: Wavelength (power)
             - 405nm (30mW)
             - 488nm (300mW)
             - 532nm (500mW)
             - 642nm (500mW)

Objectives:
             - HCX PLAPO 160x/1.43 Oil CORR GSD
             - HC PLAPO 10x/0.40
             - HCX PL FLUOTAR L 40x/0.60
             Note: Field of view with GSD objective 18x18μm or 51x51μm

Recommended Fluorophores:
             - AlexaFluor 488, 532, 546, 555, 568, 647, 680, 700
               (Life  Technologies)
             - Atto 488 520, 633, 647n, 655 (Sigma Aldrich)
             - Chromeo 488, 505 (Active Motif)
             - YFP, GFP

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.

STED can improve lateral resolution to a theoretical limit of 50nm, actual ~100nm. Axial resolution can also be improved with STED 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.

Black paneled box incubator around microscope for live cell imaging.

Lasers:
         - UV: 405nm
         - Argon: 458, 476, 488, 496, 514nm
         - White Light Laser (WLL): tunable 470nm - 670nm
         - Multi-photon 1: tunable 690nm - 1300nm
         - Multi-photon 2: 1040nm
         - STED 592nm
         - STED 775nm

Detectors:
         - Internal, AOBS: tunable 400nm - 800nm
                  - 3 standard PMTs
                  - 2 Hybrid Detectors (high sensitivity PMTs)
         - External Non-Descanned (uses filter cubes)
                  - Transmitted Light: 2 standard PMTs
                  - Reflected Light: 2 standard PMTs & 2 Hybrid Detectors

Specialists: Minh Huynh and Pam Young

   
     

Nikon A1R Advanced Confocal microscope
Confocal lasers: 403, 457, 488, 514, 560, 640nm

FLIM / FCS / FCCS lasers: 405, 488, 640nm

Detectors:
            DU4 uses Filter Blocks
                 - 2 standard PMTs
                 - 2 Galium Arsenide Phosphide (GaAsP; high sensitivity PMTs)

            Spectral Detector
            - uses prism to split emission wavelengths to discrete bandwidths

           2 FLIM / FCS / FCCS GaAsPs

Specialists: Ellie Kable and Pam Young

 
     

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

Lasers: 405, 488, 561, 640nm

Lumencor SpectraX LED: 395, 440, 470, 508, 555, 640nm

DMD: for photoactivation and photouncaging

Cameras: TIRF Monochrome - Andor iXon Ultra 888 (max 26 f/s at 1024x1024, 96f/s at 512x512)

Specialists: Minh Huynh and Pam Young
 
     

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

Light Source:
           - Lasers: 405, 488, 561, 640nm
           - DG4 White Light Source for Fura2, 340 and 380 excitation filters
           - Lumencor SpectraX LED: 395, 440, 470, 508, 555, 640nm
           - Halogen lamp and optics for Brightfield, Phase contrast, DIC

Cameras:
           - Brightfield Monochrome: DS-Qi2 (max 6 frames/second)
           - TIRF and SD Monochrome: Andor iXon Ultra 888 (max 26 f/s at
             1024x1024, 96f/s at 512x512)

Specialists: Minh Huynh and Pam Young
 
     

Nikon C2 Basic Confocal microscope
Upright Confocal microscope with motorised stage.

Lasers: 405, 488, 561, 640nm

Detectors: 3 standard PMTs using Filter Blocks - DAPI/Cy5, FITC, TRITC

Specialists: Minh Huynh and Pam Young
 
     

Nikon Ni-E Basic Fluoresence Widefield microscope
Upright microscope for mosaics and multipoint.

Light Source: Lumencor SpectraX LED - 395, 440, 470, 508, 555, 640nm

Cameras: Monochrome - DS-Qi2, Colour - DS-Fi2

Specialists: Minh Huynh and Pam Young
 
     

Leica DM6000 microscope
Basic upright widefield fluoresence microscope for mosaics using a combined monochrome/colour camera for fluorescence and histology. It uses a white light source (mercury lamp) and a set of filter cubes.

     Fliter Cubes          Ex                 Dichroic              Em
     - Ana
     - DAP                  BP 350/50      LP 400               BP 460/50
     - L5                    BP 480/40      LP 505               BP 527/30
     - RHO                 BP 546/10      LP 560               BP 585/40
     - Y5                    BP 620/60      LP 660               BP 700/75

Specialists: Minh Huynh and Pam Young
 
     

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

Specialists: Minh Huynh and Pam Young
 

LOCATED AT MADSEN BUILDING

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

Specialists: Minh Huynh and Pam Young
   
     

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

   

LOCATED AT BRAIN AND MIND CENTRE

Olympus VS120 Slide Scanner
This new state of the art slide scanner allows researchers to observe labelled tissues on slides, using 5 channel fluorescence, dark field, Differential Interference Contrast (DIC) and transmitted light modalities. Researchers can either acquire immunoflourescently labelled tissues, or typical staining procedures including DAB and H & E stained tissue.

The system has a robotic arm used in conjunction with 2 x 50 slide racks allowing up to 100 slides to be put under the microscope objective one slide at a time. Then the system autofocuses onto the tissue on the slides, recognises the edges of the tissue and scans the entire tissue, stitching the image in real time, before moving onto the next slide. It also has the capability of performing a Z-stack of up to 50um, to achieve a fully stitched 3D construction of the tissue of interest.

This instrument also has a standalone analysis computer with software allowing for detailed image viewing and analysis of the massive files which are produced from the acquisition system.

Specialist: Michael Kuligowski                                                                                                  


                                      
   

Zeiss MBF / Stereo Investigator Microscope
This inverted system features 4 objectives (10x, 20x, 40x, 63x) and a colour camera. It also has a 3 channel fluorescence system and has the capability of a heated stage and atmospheric gas control. The microscope has been specially designed with a motorised stage which is computer controlled. This, all being run by the Stereo Investigator software enables unbiased automated cell counting and quantification of area, lengths and volumes amongst others.
Also, images can be acquired for later offline analysis on another standalone system running the nuerolucida analysis software.

Specialist: Michael Kuligowski

 
   

Zeiss Axio Observer Z1 Live Cell Imaging System
This microscope has been set up in order to visualise and acquire long term (or very fast) images, to show cell migration, wound healing, cell proliferation amongst others, over time. This inverted microscopy system has a specially designed atmospheric chamber which is thermocontrolled to 37C and pumped with gasses to keep the cells we are observing live and happy for extended periods of time. The stage which the culture dish sits within is also heated and held tightly in place, to prevent any thermal drift over time. The machine is equipped with 5 Objectives (10x, 20x, 40x, 63x and 100x), 4 channel fluorescence, and transilluminated light, allowing for observation of a wide variety of cells.

 Specialist: Michael Kuligowski

 

 
   

Zeiss LSM 710 Confocal / Multiphoton Microscope
This is a custom built upright confocal microscope, designed to be used for both live animal intravital imaging (IVM) and for acquisition of tissue on slides. The instrument is fitted with objectives from 5x to 100x, including long working 20x and 40x dipping objectives specially designed to allow deep penetration into tissue (of up to 400um). The system also has a Chameleon Vision II titanium-sapphire fepto-second pulsed laser allowing for excitation between 690 and 1040nm. The confocal has 6 internal lasers (405nm, 458nm, 488nm, 514nm, 561nm and 633nm), and also a transilluminated light source, which can be acquired using either of the colour camera, 5 internal detectors, 2 non-scanning external detectors.

The stage can be totally removed, to allow for custom made animal holding stages to be fixed under the objective, allowing for live animal IVM to be undertaken.

Specialist: Michael Kuligowski