TY - CHAP
T1 - Super-resolution correlative light-electron microscopy using a click-chemistry approach for studying intracellular trafficking
AU - Andrian, Teodora
AU - Bakkum, Thomas
AU - van Elsland, Daphne M.
AU - Bos, Erik
AU - Koster, Abraham J.
AU - Albertazzi, Lorenzo
AU - van Kasteren, Sander I.
AU - Pujals, Sílvia
PY - 2021/1
Y1 - 2021/1
N2 - Correlative light and electron microscopy (CLEM) entails a group of multimodal imaging techniques that are combined to pinpoint to the location of fluorescently labeled molecules in the context of their ultrastructural cellular environment. Here we describe a detailed workflow for STORM-CLEM, in which STochastic Optical Reconstruction Microscopy (STORM), an optical super-resolution technique, is correlated with transmission electron microscopy (TEM). This protocol has the advantage that both imaging modalities have resolution at the nanoscale, bringing higher synergies on the information obtained. The sample is prepared according to the Tokuyasu method followed by click-chemistry labeling and STORM imaging. Then, after heavy metal staining, electron microscopy imaging is performed followed by correlation of the two images. The case study presented here is on intracellular pathogens, but the protocol is versatile and could potentially be applied to many types of samples.
AB - Correlative light and electron microscopy (CLEM) entails a group of multimodal imaging techniques that are combined to pinpoint to the location of fluorescently labeled molecules in the context of their ultrastructural cellular environment. Here we describe a detailed workflow for STORM-CLEM, in which STochastic Optical Reconstruction Microscopy (STORM), an optical super-resolution technique, is correlated with transmission electron microscopy (TEM). This protocol has the advantage that both imaging modalities have resolution at the nanoscale, bringing higher synergies on the information obtained. The sample is prepared according to the Tokuyasu method followed by click-chemistry labeling and STORM imaging. Then, after heavy metal staining, electron microscopy imaging is performed followed by correlation of the two images. The case study presented here is on intracellular pathogens, but the protocol is versatile and could potentially be applied to many types of samples.
KW - Click-chemistry
KW - Correlative light and electron microscopy
KW - Single molecule localization microscopy
KW - STochastic Optical Reconstruction Microscopy (STORM)
KW - Super-resolution microscopy
KW - Tokuyasu cryo-sectioning
KW - Transmission electron microscopy
UR - http://www.scopus.com/inward/record.url?scp=85092625653&partnerID=8YFLogxK
U2 - 10.1016/bs.mcb.2020.09.001
DO - 10.1016/bs.mcb.2020.09.001
M3 - Chapter
C2 - 33707017
AN - SCOPUS:85092625653
SN - 9780128220580
T3 - Methods in Cell Biology
SP - 303
EP - 331
BT - Correlative Light and Electron Microscopy IV
A2 - Müller-Reichert, Thomas
A2 - Verkade, Paul
PB - Elsevier
ER -