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ConfoCor 1 (Zeiss) Inverted confocal microscope for: - FCS measurements in solution

FCCS (Fluorescence Cross Correlation Spectroscopy). FCS (Fluorescence Correlation Spectroscopy) Technique for measuring fluorescence fluctuations in an open confocal volume (< 1 fL). Interactions and enzyme reactions in solutions or living cells can be measured in a non-destructive way.

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ConfoCor 1 (Zeiss) Inverted confocal microscope for: - FCS measurements in solution

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  1. FCCS (Fluorescence Cross Correlation Spectroscopy) FCS (Fluorescence Correlation Spectroscopy) Technique for measuring fluorescence fluctuations in an open confocal volume (< 1 fL). Interactions and enzyme reactions in solutions or living cells can be measured in a non-destructive way. FCS measurement on molecules with different fluorescent labels, which are simultaneously excited with 2 laser beams. A correlation between the two molecules such as protein-protein interactions can be measured. TCSPC (time correlated single photon counting) General Techniques • Incubators Temperature controlled growth of bacteria or eukaryotic cells (protein expression) • FPLC system Purification of proteins • Spectrofotometer Measurements of absorbance in a temperature controlled way • Calorimetry Measuring the energy of biomolecular interactions Fluorescence Techniques • ConfoCor 1 (Zeiss) Inverted confocal microscope for: - FCS measurements in solution - Fluorescence lifetime analysis - (F)RET • ConfoCor 2 (Zeiss) Inverted confocal microscope for: - FCS and FCCS in solution and cells - LSM unit - FRAP • Saffire² (Tecan) Platereader with temperature control which allows fast measurement of: - Absorbance - Fluorescence (top and bottom) - (F)RET - Polarisation • Fluorimeter (PTI) High precision fluorescence measurements - Tryptophan fluorescence - quenching studies • TCSPC setup - Temperature controlled tryptophan lifetime studies - Anisotropy measurements LSM (Laser Scanning Microscopy) Images are created by sequentially measuring all the pixels in an image. A focussed laser beam scans across the sample nd measures the intensities at each position (F)RET (Förster) Resonance Energy transfer This method measures the energy transfer between a donor and acceptor molecule. This allows to measure the interactions and distances (spectroscopic ruler) between two molecules. The emission spectrum of the donor should overlap the excitation spectrum of the acceptor. A pulsed lasersource excites fluorophores at a high frequency. The signal of the emitted photons is collected as well as the time information of the laser pulse. With this setup single photons of fluorophores are counted and correlated in time. This allows to measure the fluorescence lifetime of synthetic molecules or the natural protein fluorophore tryptophan. Tryptophan studies allow measurements on proteins in a non-destructive way.

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