Microscope Set-up

The FCS and BIFL experiments are carried out with the confocal epi-illuminated setup depicted in Fig. 1. The fluorescent molecules under study are excited by a linearly polarized, active-mode-locked argon-ion-laser (528 nm or 496 nm, 73 MHz, 150 ps). The fluorescence is detected in a confocal microscope (objective: Olympus 60x, N.A. 1.2 immersion objective; pinhole: d = 100 µm). The collected fluorescence is divided into its parallel and perpendicular components via a polarizing beamsplitter and by dichroitic beamsplitters into wavelength ranges below (Detector 1+2) and above (Detector 3+4) 620 nm. All measurements are made on freely diffusing molecules in an open detection volume (v; = 0.5 µm, z0 = 1µm, I0 = 150 kW/cm2).

Figure 1: Four-dimensional setup for detection of fluorescence parameters fluorescence intensity, F, lifetime, t, anisotropy, r, and spectral range (below and above 620 nm; filters: HQ 535/50, HQ 680/60).
One perspective of the lab
Our microscope
Verantwortlich für den Inhalt: E-Mail sendenProf. Dr. Claus Seidel