A polarized TIRFM by using a specific dye DiI

Localized topological changes of the plasma membrane upon exocytosis visualized by polarized TIRFM.
Anantharam A, Onoa B, Edwards RH, Holz RW, Axelrod D.
J Cell Biol. 2010 Feb 8;188(3):415-28.

其中的计算超过了我的理解能力

很厉害的TIRF的应用，来自一个有传统的组
貌似目前唯一的局限在DiI

Total internal reflection fluorescence microscopy (TIRFM) images the plasma membrane-cytosol interface and has allowed insights into the behavior of individual secretory granules before and during exocytosis. Much less is known about the dynamics of the other partner in exocytosis, the plasma membrane. In this study, we report the implementation of a TIRFM-based polarization technique to detect rapid submicrometer changes in plasma membrane topology as a result of exocytosis. A theoretical analysis of the technique is presented together with image simulations of predicted topologies of the postfusion granule membrane-plasma membrane complex. Experiments on diI-stained bovine adrenal chromaffin cells using polarized TIRFM demonstrate rapid and varied submicrometer changes in plasma membrane topology at sites of exocytosis that occur immediately upon fusion. We provide direct evidence for a persistent curvature in the exocytotic region that is altered by inhibition of dynamin guanosine triphosphatase activity and is temporally distinct from endocytosis measured by VMAT2-pHluorin.

[PMID: 20142424]

© 2014 En.dogeno.us. blogged by cail.cn - 35cd076bdd09fcbdfe7ca24eb8f93900