TY - JOUR
T1 - Direct Visualization and Identification of Membrane Voltage-Gated Sodium Channels from Human iPSC-Derived Neurons by Multiple Imaging and Light Enhanced Spectroscopy
AU - Moretti, Manola
AU - Limongi, Tania
AU - Testi, Claudia
AU - Milanetti, Edoardo
AU - De Angelis, Maria Teresa
AU - Parrotta, Elvira I
AU - Scalise, Stefania
AU - Santamaria, Gianluca
AU - Allione, Marco
AU - Lopatin, Sergei
AU - Torre, Bruno
AU - Zhang, Peng
AU - Marini, Monica
AU - Perozziello, Gerardo
AU - Candeloro, Patrizio
AU - Pirri, Candido Fabrizio
AU - Ruocco, Giancarlo
AU - Cuda, Giovanni
AU - Di Fabrizio, Enzo
N1 - KAUST Repository Item: Exported on 2022-05-23
Acknowledged KAUST grant number(s): OCRF-2014-CRG, OCRF-2016-CRG
Acknowledgements: The authors acknowledge financial support from King Abdullah University of Science and Technology for OCRF-2014-CRG and OCRF-2016-CRG grants; SHAHEEN cluster to provide machine time; Professor Charlotte A. E. Hauser for providing access to SHAHEEN, and from Piedmont Region through European Funds for Regional Development (“Food Digital Monitoring” project); European Research Council Synergy grant ASTRA (n. 855923).
PY - 2022/5/20
Y1 - 2022/5/20
N2 - 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 Nav 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.
AB - 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 Nav 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.
UR - http://hdl.handle.net/10754/678098
UR - https://onlinelibrary.wiley.com/doi/10.1002/smtd.202200402
U2 - 10.1002/smtd.202200402
DO - 10.1002/smtd.202200402
M3 - Article
C2 - 35595684
SN - 2366-9608
SP - 2200402
JO - Small Methods
JF - Small Methods
ER -
