Confocal laser scanning microscopy (CLSM)

The Wolfson Bioimaging Facility is exceptionally well equipped for confocal laser scanning microscopy (CLSM), the most widely used technique we provide. CLSM offers enhanced optical sectioning when compared to conventional (widefield) microscopy.

Benefits

  • Additional image clarity by removal of out-of-focus signal
  • Greater control of magnification ('zooming')
  • Specialised live-cell fluorescence techniques, eg:
    • FRAP (fluorescence recovery after photobleaching)
    • FLIP (fluorescence loss in photobleaching)
    • FRET (Förster resonance energy transfer)

Microscopes

There are six dedicated CLSM systems available. All are inverted Leica systems of varying specifications, offering a range of functionality from 'entry-level'; to more advanced systems.

Features

  • Lasers cover range of 405 - 633nm
  • Imaging of blue, green, red and far-red fluorophores
  • Phase contrast and/or DIC imaging.
  • Spectrophotometer-based detection.
  • Most have incubators for temperature control and CO2 enrichment.
  • Most have motorised stages for multi-site and 'mosaic' scanning
  • Three have additional features for live cell/organism imaging:

See our equipment list below for full details of each system.

Other CLSM systems

CLSM imaging is also available on our multiphoton and STED/FLIM/FCS systems.

  • Multiphoton:
    • Confocal imaging in upright orientation and with longer working distance objectives
  • Super-resolution microscopy/Fluorescence lifetime imaging (FLIM)/Fluorescence correlation spectroscopy (FCS):
    • Additional excitation wavelengths, 442nm and (tuneable) 470 - 670nm
    • Excitation-Emission scanning
    • Gated imaging options
    • The hybrid detectors on this system are ‘better’ than others. They are SMD HyDs for single molecule detection enabling photon counting applications like FLIM, but also have advantages for imaging as should be lowest noise.
  • Hyvolution
    • While HyVolution is not a true super-resolution technique like STED it is more broadly applicable, providing significantly improved resolution (to ~140nm) with specimens prepared for standard confocal imaging.

See our equipment list below for full details of each system.

Equipment

SystemModelSoftware modules

Laser lines

RoomFurther information
Confocal 7
Multi-laser CLSM
Leica SP5II

FRAP
FRET
Live Data Mode
Matrix Screener

405, 458, 476, 488, 496, 514, 561, 633. C29

Confocal 7 technical specifications (PDF, 183kB)

User guide (PDF, 726kB), Confocal tutorial

Confocal 8
Multi-laser CLSM - optimised for live organism imaging
Leica SP5II

FRAP
FRET
Live Data Mode

405, 458, 476, 488, 496, 514, 561, 594, 633. B9B

Confocal 8 technical specifications (PDF, 185kB)

User guide (PDF, 854kB), Confocal tutorial

Confocal 9
Multi-laser CLSM - optimised for live cell imaging
Leica SP8

FRAP
FRET
Live Data Mode
Matrix Screener

405, 458, 476, 488, 496, 514, 561, 594, 633. C35

Confocal 9 technical specifications (PDF, 187kB)

Confocal tutorial

Confocal 10
Entry level multi-laser CLSM. No motorised stage or incubator.
Leica SPE FRAP

405, 488, 532, 635.

C29

Confocal 10 technical specifications (PDF, 178kB)

Confocal tutorial

Confocal 11
Multi-laser CLSM - optimised for live organism imaging

Leica SP8   458, 476, 488, 496, 514, 561, 633. C35 Confocal 11 technical specifications (PDF, 154kB)

Confocal 12 (coming soon)
Multi-laser CLSM - optimised for live organism imaging

Leica SP8 Lightning 3D 405, 458, 476, 488, 496, 514, 561, 633 C29 Confocal 12 technical specifications coming soon

Confocal 13 (coming soon)
(Coming soon) Multi-laser CLSM - optimised for live organism imaging

Leica SP8 Lightning 3D 405, 458, 476, 488, 496, 514, 561, 633 C27A Confocal 13 technical specifications coming soon

Confocal LSB
Multi-laser CLSM located in the Life Sciences Building

Leica SP5   405, 458, 476, 488, 496, 514, 561, 633. LSB Confocal LSB technical specifications (PDF, 179kB)

Confocal LSB-2 (coming soon)
(Coming soon) Multi-laser CLSM - optimised for live organism imaging

Leica SP8 Lightning 3D 405, 458, 476, 488, 496, 514, 561, 633 LSB 326 Confocal LSB-2 technical specifications coming soon

Multiphoton

Upright CLSM/MP system with Spectra Physics DeepSee laser

Leica SP8

FRAP
FRET
Live Data Mode

458, 476, 488, 496, 514, 561, 633,

tunable 680 - 1300, fixed 1040

F4

Multiphoton technical specifications (PDF, 242kB)

User guide (PDF, 2,325kB)

STED/FLIM/FCS

CLSM with tuneable and pulsed white light laser, 592 and 660nm depletion lasers and single molecule detection

Leica SP8X

STED 
FCS 
FLIM 
HyVolution 
FRAP 
FRET

405, 442,


WLL 470 - 670,
STED 592, 660
B10B

STED/FLIM/FCS technical specifications (PDF, 179kB)

STED tutorial, FLIM tutorial, FCS tutorial

More information and access

For further information or to arrange access to this equipment, contact one of the team.

We welcome comments or suggestions. Please contact one of the team or one of the steering group.

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