Cells were transferred unto fibronectin-coated (6.5 μg/ml, Roche Diagnostics, Basel, Switzerland) transwell chambers (3.0 μm pore size, Corning Costar, Tewksburry, MA, USA) and SDF-1α (100 ng/ml) or CCL21 (300 ng/ml, both from PeproTech, Hamburg, Germany) was added.
For time-lapse microscopy, coverslips were placed in a Ludin chamber (Life Imaging Services, Basel, Switzerland) filled with imaging buffer (120 mM NaCl, 3 mM KCl, 2 mM CaCl2, 2 mM MgCl2, 10 mM glucose, 10 mM HEPES, and 2%B277, pH 7.35, 250 mOsm to match culture growth medium) (Lu et al., 2007).
Cultures of dissociated hippocampal neurons were co-infected with adenovirus on DIV7 and imaged after two days in a Ludin imaging chamber (Life Imaging Services, Basel, Switzerland) under constant perfusion with (in mM): 119 NaCl, 2.5 KCl, 2 CaCl2, 2 MgCl2, 30 glucose, and 25 HEPES, pH 7.4.
Laser scanning confocal microscopy was performed with a Leica DI6000 microscope stand connected to a SP5 scan head equipped with a temperature controlled chamber (Cube, LIS, Basel).
Equal amounts of Cy3- and Cy5-labeled cDNA was pooled and co-hybridized to a microarray slide using a EasyHyb buffer (Roche, Basel, Switzerland) in a hybridization chamber (GeneMachines) or a MAUI® mixer (BioMicro Systems, Salt Lake City, UT) at 42°C overnight.
Basal cell media (Lonza, Basel, Switzerland) supplemented with 0.5% BSA was added to the lower chamber.
The cells were incubated for 3 h at 37 °C and fluid was aspirated from the lower chamber and cell counts performed in triplicate using a Casy TT counter (Roche Innovatis, Basel, Switzerland).
Alkaline Phosphatase-conjugated anti-digoxigenin antibody (Roche Biochemicals, Basel, Switzerland) in blocking solution was then added to slides and incubated overnight at 4°C in a humidified chamber.
The Basel Chamber Orchestra provides enlightened support.
One of many notable works commissioned by the conductor and philanthropist Paul Sacher for his Basel Chamber Orchestra, it was completed in 1941 and arguably shows, in its moody and acerbic first two movements, the strains of wartime.
