Cm200 feg transmission em

For the cryo-TEM, samples of Psy 200 µM, LCN 10 nM and Psy 200 µM + LCN 10 nM were prepared in ultrapure water.
Briefly, 3 μl of sample were applied to Quantifoil holey carbon grids (copper Multi A, Quantifoil Micro Tools GmbH, Jena, Germany). Excess fluid was blotted from the grid for ~ 3 s with Whatman filter paper and then plunge frozen in liquid ethane using a home-made plunge freezer to achieve sample vitrification. Frozen samples were stored in liquid nitrogen until EM imaging. Vitrified samples were imaged using a CM200 FEG transmission EM (FEI, Eindhoven, the Netherlands) operated at 200 keV and equipped with a F224HD 2048 × 2048 CCD camera (TVIPS, Gauting, Germany). EM images were acquired at 50,000× magnification (pixel size 0.331 nm).

Morphology was observed by Cryo Electron Microscopy. 3 μL of sample were applied on Quantifoil^® holey carbon grids (copper Multi A, Quantifoil^® Micro Tools GmbH, Jena, Germany). Excess fluid was blotted from the grid for 2 sec with Whatman filter paper and then plunge frozen in liquid ethane, using a plunge freezer, to achieve sample vitrification. Frozen samples were stored in liquid nitrogen until EM imaging. Vitrified samples were imaged using a CM200 FEG transmission EM (FEI, Eindhoven, the Netherlands) operated at 200 keV and equipped with a F224HD 2048 × 2048 CCD camera (TVIPS Gauting, Germany). EM images were acquired at 27,500× magnification (pixel size 0,602 nm) at −12, −18 μm defocus.