Advanced fluorescence microscopy including single-molecule localization-based super-resolution imaging techniques requires bright and photostable dyes orproteins asfluorophores.The photophysical properties of fluorophores have been proven to be crucial for super-resolution microscopy's localization precision and imaging resolution.Fluorophores TAMRA and Atto Rho6 G,which can interact with macrocyclic host cucurbit[7]uril(CB7) to form host-guest compounds,were found to improve the fluorescence intensity and lifetimes of these dyes.We enhanced the localization precision of direct stochastic optical reconstruction microscopy(dSTORM) by introducing CB7 into the imaging buffer,and showed that the number of photons as well as localizations of both TAMRA and Atto Rho6 G increase over 2 times.
Min ZhangJing GaoJunling ChenMingjun CaiJunguang JiangZhiyuan TianHongda Wang
Cell membranes play a key role in cellular activities.Fish erythrocytes,prototype of the nucleated erythrocytes of lower vertebrates,are predominantly oval,biconvex discs with elliptical nucleus and much larger in size than human erythrocytes.In attempts to disclose the correlation of membrane structure between fish and human erythrocytes,we used in situ atomic force microscope(AFM)combined with single-molecule force spectroscopy to study the membrane structure of Crucian carp erythrocytes under quasi-native conditions.Our results revealed the asymmetric distribution of proteins in Crucian carp erythrocyte membrane:the outer leaflet of membrane is rather smooth without any proteins,whereas the inner leaflet of membrane is very rough with dense proteins.The asymmetry of fish erythrocyte membrane structure fits well with the semi-mosaic model of human erythrocyte membrane structure.This similarity of membrane structure between human and fish erythrocytes extends the semimosaic model of erythrocytes membrane structure to a wider range of species.
Yong-Mei TianMing-Jun CaiWei-Dong ZhaoShao-Wen WangQi-Wei QinHong-Da Wang
