Sample preparation for cryogenic transmission electron microscopy (cryo-TEM) was performed as in our previous work72 (link). Briefly, samples were deposited onto a 300 mesh Lacey grid (Ted Pella Ltd.), blotted and vitrified with Vitrobot Mark IV (FEI Co.). Micrographs were obtained at −177 °C using a Tecnai G2 Spirit Twin T-12 TEM (FEI Co.) at 120 kV acceleration voltage, in a low-dose mode. Images were recorded on a 4K × 4K FEI Eagle CCD camera at defocus values between 1.5 and 3 μm. Disassembly ki netic experiments were obtained by measuring the time passing between mixing wtSV40 in saline solution with carbonate buffer (CAPS buffer pH 10.7, see Section 12 in SI ) to a final concentration of 50 mM and the moment at which the grid was plunged into the liquid ethane. 30 sec was the shortest recordable time point with our setup. Capsids were characterized by their diameter. If a capsid assumed an oval shape (as in pH 10.7), its long axis was reported as its diameter.
Tecnai g2 spirit twin t 12 tem
Manufactured by Thermo Fisher Scientific
The Tecnai G2 Spirit Twin T-12 TEM is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of materials at the nanoscale. It features a twin-lens system that allows for simultaneous bright-field and dark-field imaging, providing comprehensive characterization of the sample. The instrument is capable of achieving a resolution of up to 0.24 nanometers, enabling the visualization and analysis of fine details within a wide range of specimens.
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2 protocols using tecnai g2 spirit twin t 12 tem
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Cryo-TEM sample preparation and disassembly kinetics
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Characterization of Novel Nanomaterials
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UV–vis absorption and emission spectra were recorded on a JASCO V-570 spectrometer and Varian Cary Eclipse spectrophotometer respectively. The sample fluorescence quantum yields were evaluated in comparison with coumarin 1 in ethanol as the reference (excited at 360 nm). Photo-selection excitation measurements were performed on Edinburgh Instruments FLS920 Fluorometer. XRD measurements were performed on a Phillips PW1830/40 diffractometer using the Cu Kα photons. TEM and HRTEM images were obtained on FEI Tecnai G2 Spirit Twin T-12 TEM and F20 G2 HRTEM. High-resolution HAADF STEM images and elemental mapping were obtained on FEI Themis G3 at 300 keV.
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