Exosome bound beads were resuspended in 200 μL 1× cold PBS solution and washed 2 times with pure water followed by the fixation in a 2% EMS-quality paraformaldehyde aqueous solution for electron microscopic imaging. A 5 μL sample was added to cleaned silicon chips and immobilized after air dry in a ventilation hood. Samples on silicon chips were mounted on a SEM stage by carbon paste. A ~5 nm coating of gold-palladium alloy was applied to improve SEM image background. The SEM imaging was performed under low beam energies (7 kV) on a Hitachi SU8230 filed emission scanning electron microscope. For immune staining TEM imaging, a solution of 1× PBS and 0.1% BSA (1 mL) was prepared, then a 50 μL of the solution was mixed with ~2.5 μL gold conjugated CD63 antibody (6 nM) and the prepared exosome sample grid in a clean 1.5 mL Eppendorf tube. This was incubated for 1 h at room temperature. Afterwards, the sample grid was be soaked in 100 μL MilliQ pure water for 30 sec. and transferred onto filter paper to remove all liquid and dried at room temperature for 20 min. Five minutes of negative staining with 2% aqueous uranyl acetate was followed by distilled water washes. Images were acquired at 75 kV using a Hitachi H-8100 transmission electron microscope.
Su8230 filed
Manufactured by Hitachi
The SU8230 is a high-performance scanning electron microscope (SEM) designed for a wide range of applications. It features a stable electron beam, precise sample stage control, and advanced imaging capabilities. The SU8230 provides high-resolution imaging and analysis for materials science, nanotechnology, and other research fields.
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2 protocols using su8230 filed
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Exosome Imaging via SEM and TEM
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Exosome Imaging via SEM and TEM
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Exosome bound beads were resuspended in 200 μL 1× cold PBS solution and washed 2 times with pure water followed by the fixation in a 2% EMS-quality paraformaldehyde aqueous solution for electron microscopic imaging. A 5 μL sample was added to cleaned silicon chips and immobilized after air dry in a ventilation hood. Samples on silicon chips were mounted on a SEM stage by carbon paste. A ~5 nm coating of gold-palladium alloy was applied to improve SEM image background. The SEM imaging was performed under low beam energies (7 kV) on a Hitachi SU8230 filed emission scanning electron microscope. For immune staining TEM imaging, a solution of 1× PBS and 0.1% BSA (1 mL) was prepared, then a 50 μL of the solution was mixed with ~2.5 μL gold conjugated CD63 antibody (6 nM) and the prepared exosome sample grid in a clean 1.5 mL Eppendorf tube. This was incubated for 1 h at room temperature. Afterwards, the sample grid was be soaked in 100 μL MilliQ pure water for 30 sec. and transferred onto filter paper to remove all liquid and dried at room temperature for 20 min. Five minutes of negative staining with 2% aqueous uranyl acetate was followed by distilled water washes. Images were acquired at 75 kV using a Hitachi H-8100 transmission electron microscope.
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