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Immunogold FIB‐SEM: Combining Volumetric Ultrastructure Visualization with 3D Biomolecular Analysis to Dissect Cell–Environment Interactions
Author(s) -
Gopal Sahana,
Chiappini Ciro,
Armstrong James P. K.,
Chen Qu,
Serio Andrea,
Hsu ChiaChen,
Meinert Christoph,
Klein Travis J.,
Hutmacher Dietmar W.,
Rothery Stephen,
Stevens Molly M.
Publication year - 2019
Publication title -
advanced materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 10.707
H-Index - 527
eISSN - 1521-4095
pISSN - 0935-9648
DOI - 10.1002/adma.201900488
Subject(s) - immunogold labelling , materials science , nanotechnology , nanoscopic scale , focused ion beam , ultrastructure , resolution (logic) , scanning electron microscope , in situ , biophysics , biology , chemistry , computer science , ion , organic chemistry , artificial intelligence , composite material , anatomy
Volumetric imaging techniques capable of correlating structural and functional information with nanoscale resolution are necessary to broaden the insight into cellular processes within complex biological systems. The recent emergence of focused ion beam scanning electron microscopy (FIB‐SEM) has provided unparalleled insight through the volumetric investigation of ultrastructure; however, it does not provide biomolecular information at equivalent resolution. Here, immunogold FIB‐SEM, which combines antigen labeling with in situ FIB‐SEM imaging, is developed in order to spatially map ultrastructural and biomolecular information simultaneously. This method is applied to investigate two different cell–material systems: the localization of histone epigenetic modifications in neural stem cells cultured on microstructured substrates and the distribution of nuclear pore complexes in myoblasts differentiated on a soft hydrogel surface. Immunogold FIB‐SEM offers the potential for broad applicability to correlate structure and function with nanoscale resolution when addressing questions across cell biology, biomaterials, and regenerative medicine.