摘要: The freeze-fracture technique consists of physically breaking apart (fracturing) a frozen biological sample; structural detail exposed by the fracture plane is then visualized vacuum-deposition platinum-carbon to make replica for examination in transmission electron microscope. four key steps making are (i) rapid freezing, (ii) fracturing, (iii) replication and (iv) cleaning. In routine protocols, pretreatment step carried out before typically comprising fixation glutaraldehyde followed cryoprotection with glycerol. An optional etching step, involving vacuum sublimation ice, may be after fracturing. Freeze unique among microscopic techniques providing planar views internal organization membranes. Deep ultrarapidly samples permits visualization surface structure cells their components. Images provided freeze related have profoundly shaped our understanding functional morphology cell.
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