Direct Visualization and Identification of Membrane Voltage-Gated Sodium Channels from Human iPSC-Derived Neurons by Multiple Imaging and Light Enhanced Spectroscopy.
Manola MorettiTania LimongiClaudia TestiEdoardo MilanettiMaria Teresa De AngelisElvira I ParrottaStefania ScaliseGianluca SantamariaMarco AllioneSergei LopatinBruno TorrePeng ZhangMonica MariniGerardo PerozzielloPatrizio CandeloroCandido Fabrizio PirriGiancarlo RuoccoGiovanni CudaEnzo Di FabrizioPublished in: Small methods (2022)
In this study, transmission electron microscopy atomic force microscopy, and surface enhanced Raman spectroscopy are combined through a direct imaging approach, to gather structural and chemical information of complex molecular systems such as ion channels in their original plasma membrane. Customized microfabricated sample holder allows to characterize Na v channels embedded in the original plasma membrane extracted from neuronal cells that are derived from healthy human induced pluripotent stem cells. The identification of the channels is accomplished by using two different approaches, one of them widely used in cryo-EM (the particle analysis method) and the other based on a novel Zernike Polynomial expansion of the images bitmap. This approach allows to carry out a whole series of investigations, one complementary to the other, on the same sample, preserving its state as close as possible to the original membrane configuration.
Keyphrases
- induced pluripotent stem cells
- atomic force microscopy
- high resolution
- raman spectroscopy
- electron microscopy
- endothelial cells
- single molecule
- induced apoptosis
- high speed
- spinal cord
- deep learning
- mass spectrometry
- optical coherence tomography
- healthcare
- machine learning
- convolutional neural network
- health information
- brain injury
- subarachnoid hemorrhage
- cell proliferation