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cryo fib milling  (Ted Pella)
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Ted Pella cryo fib milling
Experimental workflow imaging mitochondrial fission by cryo-ET. ( 1 ) Mammalian cells were grown on fibronectin-coated EM grids for 24–48 h and treated with FCCP for 3–6 min to induce mitochondrial fission. ( 2 ) Samples were then plunge frozen and ( 3 <t>)</t> <t>cryo-FIB</t> milled to create thin lamellae randomly positioned near the middle of the cell. ( 4 ) Lamellae were imaged by cryo-ET to create tilt series, sets of projection images obtained by iteratively tilting the sample that are used to ( 5 ) computationally reconstruct a 3D volume (a tomogram). Images in ( 3 ) are adapted from ( 22 ).
Cryo Fib Milling, supplied by Ted Pella, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cryo fib milling - by Bioz Stars, 2026-10
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Experimental workflow imaging mitochondrial fission by cryo-ET. ( 1 ) Mammalian cells were grown on fibronectin-coated EM grids for 24–48 h and treated with FCCP for 3–6 min to induce mitochondrial fission. ( 2 ) Samples were then plunge frozen and ( 3 ) cryo-FIB milled to create thin lamellae randomly positioned near the middle of the cell. ( 4 ) Lamellae were imaged by cryo-ET to create tilt series, sets of projection images obtained by iteratively tilting the sample that are used to ( 5 ) computationally reconstruct a 3D volume (a tomogram). Images in ( 3 ) are adapted from ( 22 ).

Journal: Biophysical Journal

Article Title: Nanoscale details of mitochondrial constriction revealed by cryoelectron tomography

doi: 10.1016/j.bpj.2023.07.030

Figure Lengend Snippet: Experimental workflow imaging mitochondrial fission by cryo-ET. ( 1 ) Mammalian cells were grown on fibronectin-coated EM grids for 24–48 h and treated with FCCP for 3–6 min to induce mitochondrial fission. ( 2 ) Samples were then plunge frozen and ( 3 ) cryo-FIB milled to create thin lamellae randomly positioned near the middle of the cell. ( 4 ) Lamellae were imaged by cryo-ET to create tilt series, sets of projection images obtained by iteratively tilting the sample that are used to ( 5 ) computationally reconstruct a 3D volume (a tomogram). Images in ( 3 ) are adapted from ( 22 ).

Article Snippet: In the few cases where cryo-ET was performed on peripheral cell regions without cryo-FIB milling (INS-1E cells)), Au fiducials (Ted Pella, Redding, CA, USA, cat.no.

Techniques: Imaging, Tomography