Confocal 1

Inverted Leica STED 3X Confocal with FLIM

Leica STED Confocal training manual            Location: Rm 6.029a           Bookings           Contact

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Room 6.029a - Leica DMi8 Inverted Microscope Stand with SP8 Galvo and Resonant Confocal Scanners, White Light Laser (WLL), STED 3X super-resolution, and FLIM/FRET FALCON & Lightning Deconvolution

Inverted point-scanning laser confocal microscope with spectral, super resolution and FLIM/FRET detection. Tuneable 470nm - 670nm WLL, 2 x PMT and 2 x HyD detectors. Suited for imaging fixed samples on slides, samples with low fluorescence and live imaging.

Features

  • Fully motorised X-Y-Z stage
  • High-sensitivity HyD detectors 
  • CO2 and temperature incubation for live cells and tissue
  • Tiled imaging
  • Multi-position imaging
  • Spectral imaging
  • Super-Resolution Imaging with 592nm, 660nm and 775nm STED lasers
  • Fast Resonant Imaging
  • FLIM/FRET Imaging with WLL and 440nm pulsed lasers
  • Adaptive focus control 

 

Transmitted Light:  Leica LED Lamp

Reflected Light:  Leica EL6000 120W White Light Lamp

Installed in early 2017 is the Leica STED 3X Super Resolution Microscope, which provides multicolour super-resolution imaging of cells. It is the only microscope of its type in Queensland capable of resolving the interactions between single molecules within a living cell in real time.  The system comes complete with incubation for live imaging, a resonant scan head capable of 8KHz scan speeds, the super-resolution STED module and also Fluorescent Lifetime Imaging Microscopy (FLIM) capabilities making it a truly versatile and powerful imaging system.

“The STED microscope allows us to look at cells with ultra high magnification in three dimensions. We can reconstruct cancer tissues or small animals, like fish or fly’s, in 3D. It gives us a whole new way to look at cells in health and in disease.” Professor Jenny Stow, Leader The Stow Group - Protein Trafficking and Inflammation.   

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Condensor

Name N.A. W.D. mm Position 1 Position 2 Position 3 Position 4 Position 5 Position 6
Universal LWD 0.55 28 - K3 K6 K7 - -

 

Objectives

Position Objective Magnification N.A. Immersion W.D. (mm) Type Type Thread
1 HC Plan Apochromat 10x 0.4 Dry 2.2 DIC    
2 HC Plan Apochromat 20x 0.75 Multi 0.67 DIC    
3 HC Plan Apochromat 40x 1.1 Water 0.65 DIC    
                 
5

HC Plan Apochromat(*)

93x 1.30 Glyc 0.3 DIC    
6 HC Plan Apochromat 100x 1.40 Oil 0.13 DIC    

Note: The 93x objective also comes with a motorised correction collar.

 

Fluorescent Filter Sets for Locating through the Eyepiece

Position Name Excitation Dichroic Emission Suitable Dyes
1

DAPI

350/50

 

460/50

DAPI, Hoechst

2

GFP

470/40

 

525/50

EGFP, A488

3

YFP

500/20

 

535/30

YFP

4 Texas Red 560/40   630/75 Texas Red, mCherry

 

Lasers

Laser

Wavelength

Laser Power

BFP Power*

Suitable Dyes

405

405nm

50mW

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DAPI / Hoechst

440 pulsed

440nm

-

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CFP FLIM/FRET

442

442nm 40mW - CFP
WLL

470nm-670nm

1.5mW

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Visible Dyes

92 STED

592nm

1.5W

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Star 488, Oregeon Green

660 STED

660nm

1.5W

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Star 470sx, A546

775 STED

775nm

1.5W

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SiR

*BFP = Back Focal Plane of Objective

 

Computer

HP Z640 PC - 3GHz Xeon Processor - 64GB RAM - 24GB nVidia Quadro M6000 GPU

3TB Raid Array

2 x 30" LCD Monitor

 

Software 

Leica LASX Software

Co-localisation, 3D visualisation, LAS Navigator, FRAP, FRET, FALCON FLIM and LIGHTNING Deconvolution.

 

Accessories

Incubation chamber with CO2 and Temperature control

Contacts

Dr James Springfield
Microscopy Facility Manager​
  +61 7 334 62390
  j.springfield@imb.uq.edu.au


Dr Nicholas Condon
Research Officer, Microscopy​
  +61 7 334 62042
  n.condon@imb.uq.edu.au

Dr Deborah Barkauskas
Senior Microscopist​
  +61 7 334 62042
  d.barkauskas@imb.uq.edu.au

Mailing address

ACRF Cancer Biology Imaging Facility
Institute for Molecular Bioscience
Level 6N, 306 Carmody Road,
Building 80
University of Queensland
4072, St Lucia,
Queensland, Australia