Cell Culture

CO2 incubator (ESCO), Class II biosafety cabinet (ESCO), steam autoclave (IcanClav)
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Contact: RNDr. Michal Cagalinec, PhD.

Short term culture of primary isolated left ventricular myocytes and cortical neurons and transfection using:

• Lipofectamine2000®
• adenoviruses

Main focus of the lab is analysis of dynamic remodeling of organelles and pathophysiological processes in cardiac myocytes and neurons using fluorescent indicators and proteins, where fluorescence is detected by STED, confocal and fluorescence microscopy.

To express fluorescent proteins in the myocytes and/or neurons, circular bacterial plasmids have to be transfected into the cytoplasm either by lipofection (Lipofectamine2000®) or by adenoviral transduction.

Recently we plan to express:

• MitoKikumeGR1 – photoconvertible protein (green → red) targeted to mitochondria – fusion-fission dynamics in myocytes
• Perceval – fluorescent FRET protein (CFP-YFP) – to estimate the ratio of ATP/ADP in the cell
• Chameleon D3cpv – fluorescent FRET protein (CFP-YFP) – cytoplasmic, mitochondrial and ER/SR [Ca2+] measurement