Lci plan neofluar 63 1.3 water immersion dic m27 objective

For mitochondrial fusion rate analysis, cortical neuronal cultures were transfected with mito-KikGR1 plasmid and plasmids of interest as described earlier.³⁰ (link) A laser scanning confocal microscope (LSM 510 Duo, Carl Zeiss Microscopy GmbH, Göttingen, Germany) equipped with a LCI Plan-Neofluar 63×/1.3 water immersion DIC M27 objective (Carl Zeiss Microscopy GmbH, Göttingen, Germany) was used. The temperature was maintained at 37 °C using a climate chamber. For fusion acquisition, mito-KikGR1 was illuminated with a 488 nm argon laser line to visualize the intense green mitochondrial staining. Selected mitochondria were then photoconverted to red using a 405 nm diode laser and illuminated using a 561 nm DPSS laser. The images were taken at 10 s intervals for 10 min, the fate of all activated mitochondria was followed throughout the time-lapse, and the fusion and fission events were recorded. In experiments that compared different conditions, 4 fields per dish were imaged and at least 4 dishes per condition were used. Mitochondrial length measurements were performed as described earlier.³⁰ (link)