000201405 001__ 201405
000201405 005__ 20190317000002.0
000201405 0247_ $$2doi$$a10.1038/nprot.2014.101
000201405 022__ $$a1754-2189
000201405 02470 $$2ISI$$a000337145800012
000201405 037__ $$aARTICLE
000201405 245__ $$aSemiautomated correlative 3D electron microscopy of in vivo-imaged axons and dendrites
000201405 260__ $$bNature Publishing Group$$c2014$$aLondon
000201405 269__ $$a2014
000201405 300__ $$a13
000201405 336__ $$aJournal Articles
000201405 520__ $$aThis protocol describes how in vivo-imaged dendrites and axons in adult mouse brains can subsequently be prepared and imaged with focused ion beam scanning electron microscopy (FIBSESEM). The procedure starts after in vivo imaging with chemical fixation, followed by the identification of the fluorescent structures of interest. Their position is then highlighted in the fixed tissue by burning fiducial marks with the two-photon laser. Once the section has been stained and resin-embedded, a small block is trimmed close to these marks. Serially aligned EM images are acquired through this region, using FIBSESEM, and the neurites of interest are then reconstructed semiautomatically by using the ilastik software (http://ilastik.org/). This reliable imaging and reconstruction technique avoids the use of specific labels to identify the structures of interest in the electron microscope, enabling optimal chemical fixation techniques to be applied and providing the best possible structural preservation for 3D analysis. The entire protocol takes similar to 4 d .
000201405 700__ $$0245195$$g209404$$uEcole Polytech Fed Lausanne, Ctr Electron Microscopy, BioEM Facil, CH-1015 Lausanne, Switzerland$$aMaco, Bohumil
000201405 700__ $$0240852$$g120862$$uEcole Polytech Fed Lausanne, Ctr Electron Microscopy, BioEM Facil, CH-1015 Lausanne, Switzerland$$aCantoni, Marco
000201405 700__ $$uUniv Geneva, Univ Med Ctr CMU, Dept Neurosci, Geneva, Switzerland$$aHoltmaat, Anthony
000201405 700__ $$aKreshuk, Anna
000201405 700__ $$aHamprecht, Fred A.
000201405 700__ $$aKnott, Graham W.$$uEcole Polytech Fed Lausanne, Ctr Electron Microscopy, BioEM Facil, CH-1015 Lausanne, Switzerland$$g159872$$0240043
000201405 773__ $$j9$$tNature Protocols$$k6$$q1354-1366
000201405 909C0 $$xU10659$$0252087$$pCVLAB
000201405 909C0 $$0252025$$pCIME$$xU10192
000201405 909CO $$qGLOBAL_SET$$pSB$$pIC$$particle$$ooai:infoscience.tind.io:201405
000201405 917Z8 $$x112366
000201405 937__ $$aEPFL-ARTICLE-201405
000201405 973__ $$rREVIEWED$$sPUBLISHED$$aEPFL
000201405 980__ $$aARTICLE