TEM and immuno-TEM analysis of EVs were performed as described previously [4 ] with modifications. Specifically, the vesicle was re-suspended in 1× PBS and deposited onto a formvar-coated slide of copper mesh EM grids (FF300-Cu-50, Electron Microscopy Sciences). The vesicle-coated grids were washed, stained with 1% UO2(CH3COO)2 and then viewed by TEM using a Philips CM12 with Gatan model 791 camera; for the immuno-gold labelling with antibodies, blocked grids coated with EVs were briefly fixed by ice-cold 1:1 ethanol/methanol for 5 min followed by washing with PBS. The grids were then transferred to a drop of the anti-CD9 antibody (Abcam) in PBS with 1% BSA, 0.01% Tween20 and incubated for 30 min. The grids were then washed and incubated with gold-labelled secondary antibody (Sigma). The grids were stained with 0.5% UO2(CH3COO)2 and then imaged by TEM.
Ff300 cu 50
Manufactured by Electron Microscopy Sciences
The FF300-Cu-50 is a copper grid designed for use with electron microscopy. It features a mesh size of 300 lines per inch and a grid thickness of 50 micrometers. The grid is made of copper and is suitable for a variety of electron microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ff300 cu 50
1
Visualization of Extracellular Vesicles
2
Electron Microscopy of Vesicles
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EM and immuno-EM analysis of vesicles were performed as described previously37 (link) with minor modifications. Specifically, the vesicle pellet was resuspended in 1X PBS and deposited onto a Formvar coated slide of copper mesh EM grids (FF300-Cu-50, Electron Microscopy Sciences). The vesicle-coated grids were washed, stained with 1% UO2(CH3COO)2 and then viewed for transmission EM (TEM) using a Philips CM12; camera: Gatan model 791. For the immuno-gold labelling with antibodies, blocked grids were transferred to a drop of the antibody in 0.1 M NaPO4 pH 7.2/150 mM NaCl/1% BSA/0.01% Tween20 and incubated for 30 min. The grids were then washed, incubated with gold-labeled secondary antibody in 1X PBS/1% BSA/0.01% Tween20 for 20 min. After washing with the same buffer and deionized water, the grids were stained with 0.5% UO2(CH3COO)2 and then imaged by TEM.
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