Confocal microscopes

Leica STELLARIS Confocal microscope

• white light laser providing a continuous choice of excitation wavelengths  into NIR (485-790 nm)
• 405 nm CW laser
• 448 nm CW laser
• 5 x “Power HyD” GaAsp detectors, allowing up to 5 channel simultaneous imaging when used with additional NIR capability
• “super-resolution” confocal to 120 nm resolution.
   

Objectives:

• 10x, 0.4, N.A.
• 20x, 0.75, N.A.
• 40x, 1.25, N.A., Glycerol with motorised correction collar
• 63x, 1.4, N.A., Oil

“TauSense” uses fluorescence lifetime detection for additional fluorophore separation, or distinct gating of overlapping signals into separate channels dependent on their lifetime, for example autofluorescence.

Changes in a fluorophores nano-environment can also be measured using fluorescence lifetime e.g. FRET of interacting molecules, pH etc.

Features

• transmitted light
• DIC
• hardware autofocus
• simultaneous fluorescence acquisition
• sequential fluorescence acquisition 
• mark and find
• motorised stage with Tile Scan (stitch multiple FOV together)
• time-lapse
• photo-kinetics
• FLIM
• NIR Imaging
• lambda-scan/spectral channel un-mixing
   

2 x Leica SP8 AOBS Laser Scanning Confocal Microscopes

Modality and resolution

• CLSM - 200 nm
• DIC - 250 nm

Objectives

Magnification, numerical aperture and immersion

• 10x, 0.5, air
• 20x, 0.7, air
• 40x, 1.3, oil
• 63x, 1.3, glycerol

Specimen format

• standard glass slide
• 35 mm coverslip bottom dish
• coverslip bottom chamber-slide
• optical bottom multi-well plate

Detectors

• confocal 1: 2 x PMT (photomultiplier tube), 1 x “HyD” GaAsP detector
• confocal 2: 1 x PMT, 2 x “HyD” GaAsP detector

Lasers/filters

Excitation laser λ

• 405 nm  (nano meters)  
• 458 nm    
• 476 nm    
• 488 nm    
• 496 nm    
• 514 nm    
• 561 nm    
• 594 nm    
• 633 nm   

Emission filters

• 3 x Spectral Detectors. Any λ/Bandwidth from 400-800 nm. 

Fluorescence acquisition

• simultaneous fluorescence acquisition of up to 3 channels
• sequential fluorescence acquisition is unlimited

Other features

• transmitted light
• differential interference contrast (DIC)
• time lapse
• photo-kinetics
• 3D imaging

3D specimen penetration is less than 100 μm.

The microscope does not have hardware autofocus. It has environmental temperature and gas control and a motorised stage with tile scan (stitch multiple FOV together).

It also offers mark and find and lambda-scan and spectral channel un-mixing.