Gottfried Schatz Research Center | Molecular Biology and Biochemistry

Image 1

#I-1: 3D reconstruction of mitochondria in living human endothelial cells that were transiently tansfected with ratiometric-pericam-mt (a kind gift from Prof. Atsuhi Miyawaki, Japan).

Image 2

#I-2: 3D reconstruction of mitochondria in living human endothelial cells that were transiently tansfected with mitochondria-targeted DsRed 2 (a kind gift from Prof. Tullio Pozzan, Italy) and ApoK1er (Osibow et al).

Image 3

#I-3: 3D reconstruction of the ER in living human endothelial cells that were transiently transfected with Cam4er (a kind gift from Profs. Roger Y. Tsien and Atsuhi Miyawaki)

Image 4

#I-4: High D-glucose but not high mannitol yield fragmentation of mitochondria within 2 h of incubation. Three-dimensional architecture of the mitochondrial network was monitored using an ACLSM using a 100x objective (NA 1.45) and a z-step size of 100 nm. After image deconvolution 3D restoration and surface rendering were performed using Bitplane Imaris 3.3.

Movie 1

 

#M-1: This is the way how we position the patch pipette on cell membrane that is close to superficial intracellular organelles. Mostly we apply our Physiological Patch technique (Malli et al).

Movie 2

 

#M-2: Endothellial cells were transiently transfected with cytosolic GFP and mitochondria-targeted DsRed. Cytosolic surface and mitochondrial structure are calculated by surface- and volume-rendering, respectively.

Movie 3

 

#M-3: 4D movie on the fragmentation of mitochondrial network in endothelial cells upon exposure to superoxide anions. The time frame is overall 1h and grid distance indicates 1 µm.

Movie 4

 

#M-4: Spike-like accumulation of PKCalpha in endothelial cells upon cell stimulation under conditions of distrubed intracellular Ca2+ cycling. Endothelial cells expressing PKCalpha-GFP were stimulated with BHQ & histamine and the accumulation of PKCalpha was monitored using the ACLSM.

Movie 5

 

#M-5: Mitochondrial movements are sensitive to cytosolic Ca2+ elevation. Human endothelial cells were transiently transfected with ratiometric-pericam-mt and stimulated with histamine as indicated.

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