Multicolour Confocal Laser Scanning
02 Mar 2010



Multicolour Confocal Microscopy

OCTOPUS offers several confocal microscopes to meet complex user requirements. The confocal instruments include: three Nikon scanning units (eC1 and eC2 coupled to TE2000, Ti and Ti_E), Leica SP8 (with STED) and custom built systems for unique specifications that are not commercially available. Two of the Nikon systems are also converted for multiphoton microscopy. The conversions also allows for UV imaging as well as full emission spectra at each pixel.

Our talented instrument and research scientific continually seek to develop several revolutionary imaging methods, several of which may take five to ten years to complete. With this in mind our vision is to develop the next generation advanced scanning imaging platforms with constant horizon scanning. Working closely with the ultra-fast group (ULTRA) provides us with the opportunity to combine new and emerging laser light sources with unique properties for use in our confocal microscopes.
The confocal lasers available are UV to near infrared (Ti:Sapphire laser system), continuous wave (CW) and pulse widths of nanosecond, picosecond and femtosecond. Standard excitation laser lines include: 405 nm, 488 nm, 543 nm, 561 nm, 633 nm, 641 nm as well as super continuum white light sources (Fianum and NKT) fur further flexibility.
The confocal microscopes also offer time-correlated single photon counting set up (TC-SPC). The technique allows excited state fluorescence lifetime measurements of chemicals in solution and solid samples down to a few picoseconds. The resolution of our set up (instrument response function) is 35 ps. Our primary research interests include Fluorescence Lifetime Imaging Microscopy (FLIM).
The combined confocal laser scanning microscopy with the TC-SPC technique enables us to generate 3D confocal images as well as picosecond lifetime measurements. As well as point scanning confocal microscopy, we have also developed a line scanning FLIM. This technique allows FLIM measurements at video rates when the technique is combined with an ultra-fast gated image intensifier (Andor ICCD).
Microscope specification
Instrument Nikon EC1, EC2, Leica SP8, Custom with GSI Lumonics Galvos
Laser wavelengths 405, 488, 543, 561, 642 nm and white light
Compatible objective lenses 4x, 10x, 40x air, 20x NA 0.75, 63x oil immersion, 60x water immersion NA 1.2, 60x air ELWD, 100x oil immersion NA 1.49
Compatible dyes Any
Compatible sample mounts Cover slip on microscope slide or glass bottom Petri dish
Configuration Inverted microscope
Temperature control Environmental chamber or heated stage insert, 5-50 °C. gases include N2, CO2, O2, Ar or any mixture
Lateral resolution ~250 nm (depends on wavelength)
Axial resolution ~500 nm (depends on wavelength)
  • Mammalian and plant cell biology
  • Protein-protein interactions
  • Time-lapse imaging of cellular processes
  • Biological research
  • Medical research
  • Drug discovery
  • Material science
  • Geological samples
Contact: Botchway, Stanley (STFC,RAL,CLF)