RT Journal Article
SR Electronic
T1 Correlative fluorescence microscopy, transmission electron microscopy and secondary ion mass spectrometry (CLEM-SIMS) for cellular imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.10.05.326074
DO 10.1101/2020.10.05.326074
A1 Felix Lange
A1 Paola Agüi-Gonzalez
A1 Dietmar Riedel
A1 Nhu T.N. Phan
A1 Stefan Jakobs
A1 Silvio O. Rizzoli
YR 2020
UL http://biorxiv.org/content/early/2020/10/05/2020.10.05.326074.abstract
AB Electron microscopy (EM) has been employed for decades to analyze cell structure. To also analyze the positions and functions of specific proteins, one typically relies on immuno-EM or on a correlation with fluorescence microscopy, in the form of correlated light and electron microscopy (CLEM). Nevertheless, neither of these procedures is able to also address the isotopic composition of cells. To solve this, a correlation with secondary ion mass spectrometry (SIMS) would be necessary. SIMS has been correlated in the past to EM or to fluorescence microscopy in biological samples, but not to CLEM. We achieved this here, using a protocol based on transmission EM, conventional epifluorescence microscopy and nanoSIMS. The protocol is easily applied, and enables the use of all three technologies at high performance parameters. We suggest that CLEM-SIMS will provide substantial information that is currently beyond the scope of conventional correlative approaches.