Cells were allowed to load MitoTracker Red CMXRos (#M7512) or MitoTracker Green (M7514) (both from Thermo Fisher Scientific) for 10 min, washed two times with Hank’s BSS containing calcium and magnesium, then fixed, counter-stained with Hoechst 33258 and mounted in Fluorsave (Calbiochem, Milan, Italy). MitoTracker Green was used to detect the total amount of mitochondria in cells, as it localizes to the organelle regardless of mitochondrial membrane potential. MitoTracker Red CMXRos was used to visualize only functional mitochondria, as its accumulation depends on membrane potential. Images were acquired using a ZEISS LSM 510 META confocal laser scanning microscope and analyzed by the LSM 5 software, version 3.5 (Carl Zeiss, Gottingen, Germany).
Lsm 5 software version 3
Manufactured by Zeiss
Sourced in Germany
The LSM 5 software, version 3.5, is a computer program designed for the operation and analysis of data from Zeiss microscopy systems. It provides a user interface for controlling the microscope, capturing images, and processing the acquired data.
Automatically generated - may contain errors
2 protocols using lsm 5 software version 3
1
Mitochondrial Membrane Potential Imaging
2
Visualizing Chloroplast Morphology
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CM was conducted with an LSM5 laser scanning confocal microscope equipped with an argon-krypton laser (Carl Zeiss Inc.). A 488 nm excitation line and an AOBS filter-free system were employed to collect emitted light between 498 and 700 nm. The autofluorescence of chlorophyll was used to visualize chloroplast structure. Collections of optical sections through chloroplasts were used to obtain a three-dimensional reconstruction of chloroplast morphology. The reconstructions of chloroplasts were made by LSM 5 software version 3.5 (Carl Zeiss Inc.).
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