Immunogold FIB-SEM: Combining Volumetric Ultrastructure Visualization with 3D Biomolecular Analysis to Dissect Cell–Environment Interactions

TY - JOUR

T1 - Immunogold FIB-SEM

T2 - Combining Volumetric Ultrastructure Visualization with 3D Biomolecular Analysis to Dissect Cell–Environment Interactions

AU - Gopal, Sahana

AU - Chiappini, Ciro

AU - Armstrong, James P.K.

AU - Chen, Qu

AU - Serio, Andrea

AU - Hsu, Chia Chen

AU - Meinert, Christoph

AU - Klein, Travis J.

AU - Hutmacher, Dietmar W.

AU - Rothery, Stephen

AU - Stevens, Molly M.

PY - 2019/8/9

Y1 - 2019/8/9

N2 - 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.

AB - 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.

KW - 3D reconstruction

KW - electron microscopy

KW - FIB-SEM

KW - image analysis

KW - immunogold

UR - http://www.scopus.com/inward/record.url?scp=85067403326&partnerID=8YFLogxK

U2 - 10.1002/adma.201900488

DO - 10.1002/adma.201900488

M3 - Article

AN - SCOPUS:85067403326

SN - 0935-9648

VL - 31

JO - Advanced Materials

JF - Advanced Materials

IS - 32

M1 - 1900488

ER -