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Laboratory Instrumentation in CQFM


Time-resolved fluorescence - Single photon timing

Time-resolved

                          instruments

The facility for measurements of decay times of fluorescence and fluorescence anisotropy is a home-built set-up with mode-locked laser excitation and time-correlated photon counting detection. The equipment is placed on a floating optical table in a controlled temperature room. The system includes two pump lasers, a mode-locked titanium:sapphire laser, a pulse selector, two ultrafast dye lasers, a microchannel plate-photomultiplier tube (MCP-PMT), a Single photon timing (SPT) system and a monochromator.

Excitation lasers:
  • Spectra-Physics Tsunami mode locked Ti:Sapphire femtosecond laser with continuously tuneable output emission over a broad range of near IR wavelengths (λ=700-1000nm, Imax=1.8W, t=100fs, f=80MHz). Emission at the second (λ=350-470nm) and third (λ=240-300nm) harmonics of laser is also used for sample excitation.
  • Coherent Radiation Dye laser 700 series (λ=560-680nm, IMax=130mW, f=4MHz, t=5ps, with pumping by 2W Vanguard) is used for sample excitation at first and second harmonics of Rhodamine 6G (λ=560-610nm and λ=280-305nm) or DCM (λ=610-680nm and λ=305-340nm) dye laser. It includes cavity dumper, driver and dye circulator.

Steady-state fluorescence

Horiba Jobin Yvon Fluorolog 3-22 Spectrofluorimeter

Fluorolog 3-22 Spectrofluorimeter

Measurement of excitation and emission fluorescence and phosphorescence spectra (including anisotropy) and determination of lifetimes longer than 1 microsecond. Equipped with a thermostated sample holder, a solid sample holder, a Dewar for phosphorescence measurements, a solid state IR detector (800-1550 nm), a flash lamp and polarizers. Samples can be measured at different temperatures in the -190 to 100 ºC range.

SLM Aminco 8100 Spectrofluorimeter

SLM Aminco 8100 Spectrofluorimeter

Performs steady-state fluorescence measurements. Includes the following: 450 W lamp; double excitation and emission monochromators; Glan-Thompson polarisers for automated steady-state fluorescence anisotropy measurements; thermostated sample holder with a Julabo F25 water bath circulation; Bio-Logic SFM-20 mixing device for stopped-flow kinetics.

Spex Fluorolog F112A Spectrofluorimeter

Spex Fluorolog F112A Spectrofluorimeter

Measurement of excitation and emission fluorescence and phosphorescence spectra (including anisotropy) and determination of lifetimes longer than 1 ms. Equipped with a thermostated sample holder, a solid sample holder, a Dewar for phosphorescence measurements and polarizers. Samples can be measured at different temperatures in the -190 to 100 ºC range.

 
UV-Vis absorption spectroscopy

Shimadzu UV-3101PC UV-Vis-NIR Spectrophotometer

Shimadzu UV-3101PC UV-vis-NIR

                        Spectrophotometer

Absorption spectra in the wavelength range 190 to 3200 nm. Equipped with polarizers, an integrating sphere for measurement of opaque solid materials, and a thermostated liquid sample holder (-20 to 90 ºC).

Jasco V-560 UV-Vis Spectrophotometer

Jasco V-560 UV-vis Spectrophotometer

Double-beam spectrophotometer with a photomultiplier tube detector for measurement of UV-vis (190 to 900 nm) absorption spectra.

Jasco V-660 UV-Vis Spectrophotometer

Double-beam spectrophotometer with a photomultiplier tube detector for measurement of UV-vis (190 to 900 nm) absorption spectra. Extended photometric linearity up to 6 Abs enables measurement of highly absorbing samples and is also effective for filter measurements approaching 0 %T.

Confocal Microscopy

Leica TCS-SP5 Multiphoton/Confocal Fluorescence Microscope

Leica TCS-SP5 Multiphoton/Confocal

                        Fluorescence Microscope

The Leica TCS confocal microscope allows the examination of fluorescent specimens with clarity and resolution that far exceeds that of conventional fluorescence microscopy. The fluorophrores are excited in a very narrow plane using laser illumination. The stimulated emissions are also collected from that narrow plane using sensitive photomultiplier tubes that detect light of only the selected wavelengths. The Leica software allows image manipulations for 3-D reconstruction of optical sections, creation of movies showing rotation of images, and sophisticated calculations of intensities and ratios.
Available techniques are: Imaging (2D and 3D) with simultaneous detection of up to two colors, Förster Resonant Energy Transfer imaging (FRET), Fluorescence Recovery After Photobleaching (FRAP), Fluorescence Loss in Photobleaching (FLIP), Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Lifetime Imaging (FLIM).
Accessories include: CW lasers: Ar ion (458, 465, 488, 496 and 514 nm) and HeNe (633 nm) Pulsed lasers: Ti:Saphire (710-990 nm, 100 fs, 80 MHz); Autocorrelator and two APDs set for single molecule FCS; TCSPC module coupled to a DCC-100 PMT for FLIM.

Atomic Force Microscopy

Nanosurf® easyScan 2 FlexAFM System

Nanosurf® easyScan 2 FlexAFM System

Nanosurf® easyScan 2 FlexAFM System for AFM measurements in air and liquid with active vibration isolation and micrometer translation stage. Video camera for view of sample and cantilever with simultaneous top and side view. Adapter plates for inverted microscope (for use with confocal, multiphoton and fluorescence lifetime imaging microscopy, and fluorescence correlation microscopy). Sample heating for materials research and biological samples, noise control encasing and environmental control chamber.

Other equipment

  • Cell culture room
  • BMG Labtec® POLARstar Optima multimode microplate reader                                                                                                                                           (Absorbance, Luminescence, Fluorescence Intensity, Time-resolved Fluorescence, Fluorescence Polariztion)
  • Refrigerated bench centrifuges
  • Avanti Polar Lipids® Mini-extruder
  • Dry block heaters
  • Branson® Sonifier 250 with microprobe