Instruments, computer software and materials
Microscopes and Spectometers
Olympus XL-71 Confocal FV-300 Microscope
The Olympus IX71 inverted microscope is equipped with a FluoView 300 confocal scan-head for multi-and single-photon excitation. This microscope system is used for fluorescence lifetime imaging (FLIM), fluorescence correlation spectroscopy (FCS) and single-molecule Förster resonance energy transfer (single-molecule-FRET) experiments. Perkin-Elmer Avalanche Photodiodes (APDs) are used for high-sensitivity detection.
Nikon 2000TE Microscope for Imaging and Spectroscopy
The Nikon TE2000U inverted microscope has four ports for observation and has been configured for direct excitation fluorescence imaging and Total Internal Reflection Fluorescence microscopy (TIRF) using a Cascade 512F CCD camera. The TIRF configuration currently supports single-molecule Förster resonance energy transfer (single-molecule-FRET) experiments.
FLASC 1000 Time-Correlated Single-Photon Counting Fluorometer
The FLASC 1000 Time-Correlated Single-Photon Counting (TCSPC) Fluorometer is for time-resolved fluorescence experiments. The fully automated sample chamber, designed and constructed in collaboration with Quantum Northwest (Liberty Lake, WA), uses a modified ‘T-format’ optical arrangement that permits simultaneous collection of multiple decay curves. On one side, a standard polarizer is used to collect fluorescence decays at any angle, including magic angle. On the other side, a beam-splitting polarizer is used to separate the vertical and horizontal components of the emission. Three separate emission trains separate polarization components. Each uses monochromators or filters for wavelength selection followed by separate detectors. Each of the three emission detectors has dedicated processing electronics. The sample chamber is temperature controlled and can accommodate different sample holders, including standard cuvettes, a high-pressure cell, or a goniometer for crystal samples.
SpectraMax M2 Steady-State Absorption Spectrometer & Fluorometer
The SpectraMax M2 is a dual-monochromator, multi-detection microplate reader with a dual-mode cuvette port and 6-384 microplate reading capability. This instrument performs steady state absorbance measurements with a wavelength range of 200-1000 nm, fluorescence excitation measurements from 250-850 nm, and fluorescence emission measurements in the wavelength range of 360-850 nm.
Coherent Mira Ti:Sapphire Ultrafast Laser
The Mira laser system, which consists of a 10 W diode laser (Verdi V-10) pumping a Ti:Sapphire laser (Mira 900), can be mode-locked to provide picosecond or femtosecond pulses of light and is tunable to wavelengths between 680 and 1050 nm. The Mira is used in combination with a Coherent Pulse Picker, which modifies the pulse frequency from 9kHz to 4MHz, and an InRad Harmonic Generator, which can frequency double or triple the Mira’s output to create wavelengths between 260 and 520 nm. This entire system is used as a light source for the time-resolved fluorometer as well as for the Nikon and Olympus microscopes.
Coherent Chameleon Femtosecond Laser
This Ti:Sapphire femtosecond pulsed laser (Chameleon) is dedicated to two- and three-photon microscopy experiments.
PicoQuant LDH-470, LDH-480, and LDH-635 Picosecond Diode Lasers
These fiber-coupled, picosecond-pulsed laser diodes at 470 nm, 478 nm and 637 nm, with a frequency range from single shot to 40 MHz, are used with the time-resolved fluorometer as well as the Nikon and Olympus microscopes.
Coherent OBIS 552 LS
This CW diode laser at 552 nm is used for TIRF microscopy with the Nikon TE 2000.
PicoQuant FluoFit (reconvolution lifetime and time-resolved anisotropy)
PicoQuant SymphoTime (FCS, FLIM, FRET)
OriginLab (general data fitting)