Deconvolution Microscopes

Resource Links:

• Nikon Filter Chart
• Training Videos
• Microscope location - Room 6.025c
• Make a Booking

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Room 6.025c - Nikon Ti-E Inverted Microscope Stand with Hamamatsu Flash 4.0 sCMOS camera

Inverted widefield microscope with Lumencor 7 line LED lightsource, high speed filter wheels and CMOS camera. Suited for imaging fixed samples on slides, samples with low fluorescence and live imaging.

Features Include:

• Fully motorised X-Y-Z stage
• Piezo Z-drive for rapid Z-stacks
• CO2 and temperature incubation for live cells and tissue
• Tiled imaging
• Multi-position imaging
• Transmitted Light:  Sutter 100W LED Lamp

Reflected Light:  Lumencor Spectra LED Lightsource

Condensor:

Objectives:

Fluorescent Filters:

LEDs:

*BFP = Back Focal Plane of Objective

Computer:

Dell PC - 3GHz Xeon Processor - 32GB RAM - 1GB nVidia Quadro GPU

3TB Raid Array

30" LCD Monitor

Software: 

NIS Elements 4.3

Accessories:

Incubation chamber with CO2 and Temperature control

• OBS Training Manual
• OBS Filter Chart
• Microscope location - Room 6.018
• Make a Booking

Room 6.018 - Olympus IX81 Inverted Microscope Stand with Hamamatsu Orca CCD camera

Inverted widefield microscope with Xenon lightsource, high speed filter wheels and CCD camera. Suited for imaging fixed samples on slides, samples with low fluorescence and live imaging.

Features Include:

• Fully motorised X-Y-Z stage
• CO2 and temperature incubation for live cells and tissue
• Tiled imaging
• Multi-position imaging

Transmitted Light:  Olympus 100W Halogen Lamp

Reflected Light:  Olympus MT20 Xenon White Lightsource

Condensor:

Objectives:

Fluorescent Filter Sets:

Computer:

Dell PC - 3GHz Xeon Processor - 8GB RAM - 256GB nVidia Quadro GPU

1TB Raid Array

30" LCD Monitor

Software: 

Cell^R

Accessories:

Incubation chamber with CO2 and Temperature control

Room 6.019B

The Lattice Lightsheet microscope is an advanced live cell microscope developed by Nobel Prize winner Eric Betzig in 2014. This microscope is extremely gentle on cells allowing for very long acquisitions, as well as being super-fast, (capturing full 3D volumes in under a second).

Highlights

• Very High spatial resolution (230 nm, 230 nm, 300 nm, xyz)
• Very High Temporal resolution (100 slices / second)
• Very low photo-toxicity
• Opportunity to generate very rare data (very few working systems world wide)
• 4 Individual lasers (440 nm (CFP), 488 nm (GFP), 560 nm (RFP), 640 nm (Far Red))

Challenges

• Massive data sets are generated by this system (terabytes)
• Visualisation and analysis software solutions are not equipped for large volumes of LLSM data, instead MIPS are generated for indexing of files, and finding sections of movies for further analysis. Large collaborations are underway to develop tools to fix this.
• This system is not a ‘turnkey solution’ like our Zeiss systems, therefore the software is not as refined, and often many alignments are required to optimise the imaging output.

Sample Preparation

• This microscope takes 5 mm coverslips (thickness is not important) mounted on the holder shown below.
• The stage can be moved within a 3 x 3 mm region from the centre of the coverslip
• Samples must be adhered to the coverslip (for non-single cell imaging please consult with facility staff)
• Samples are imaged with water dipping lens’ with your sample in a 8 mL water bath (take this into account if your require stimulants/drugs in your imaging media)
• The stage/water-bath is heated, however CO₂ is not available while imaging.
• The LLSM room has an incubator set to 37 ^oC 5% CO₂.