A system for optical high resolution screening of electrical excitable cells.
The application of primary excitable cells for high content screening (HCS) requires a multitude of novel developments including cell culture and multi-well plates. Here we introduce a novel system combining optimised culture conditions of primary adult cardiomyocytes with the particular needs of excitable cells for arbitrary field stimulation of individual wells. The major advancements of our design were tested in calcium imaging experiments and comprise (i) each well of the plate can be subjected to individual pulse protocols, (ii) the software driving electrical stimulation can run as a stand-alone application but also as a plug-in in HCS software packages, (iii) the optical properties of the plastic substrate (foil) resemble those of glass coverslips fostering high resolution immersion-based microscopy, (iv) the bottom of the foil is coated with an oleophobic layer that prevents immersion oil from sticking, (v) the top of the foil is coated with an elastic film. The latter enables cardiomyocytes to display loaded contractions by mimicking the physiologically occurring local elastic network (e.g. extracellular matrix) and results in significantly increased contractions (with identical calcium transients) when compared to non-elastic substrates. Thus, our novel design and culture conditions represent an essential further step towards the application of primary cultured adult cardiomyocytes for HCS applications.
Müller
Oliver
O
Institute for Molecular Cell Biology, Saarland University, Medical Faculty, Building 61, 66421 Homburg/Saar, Germany.
Tian
Qinghai
Q
Zantl
Roman
R
Kahl
Valentin
V
Lipp
Peter
P
Kaestner
Lars
L
D000200:Action Potentials / Q000502:physiology*; D000818:Animals; D020013:Calcium Signaling / Q000502:physiology*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004548:Elasticity / Q000502:physiology; D004558:Electric Stimulation / Q000295:instrumentation / Q000379:methods; D004566:Electrodes; D004582:Electronics, Medical; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods*; D005109:Extracellular Matrix / Q000502:physiology; D005456:Fluorescent Dyes; D008297:Male; D008856:Microscopy, Fluorescence; D009119:Muscle Contraction / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar; D012984:Software; D056969:Voltage-Sensitive Dye Imaging / Q000295:instrumentation / Q000379:methods*
D005456:Fluorescent Dyes