Ratiometric sensing using dual-frequency lifetime discrimination.
Ratiometric sensing using dual-frequency lifetime discrimination.
复制标题
使用双频寿命辨别的比例传感。
DOI：
10.1006/abio.2001.5336
复制
发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Rao,G
中科院分区：
文献类型：
--
作者：
Kostov,Y;Harms,P;Rao,G
Materials and methods. Three different fluorophores were used: fluorescein (Sigma, St. Louis, MO), Oregon green–BAPTA (Molecular Probes, Eugene, OR), and partially purified green fluorescent protein (GFP), 2 produced from pBAD–GFP-transformed Escherichia coli strain JM105 (15). The pH determinations were performed using phosphate buffers. The concentrations of the fluorophores were as follows: fluorescein—0.8 M, calcium-sensitive dye—0.635 M, and GFP—0.8 M. Calcium determinations were performed using calcium calibration kit No. 2 (Molecular Probes).The reference consisted of 1%(w/w) ruthenium tris-(bipyridyl) dichloride physically entrapped in cellulose acetate film by dissolving both in acetone and spread casting the mixture. As Ru (bpy) 3Cl2 is water soluble, it was necessary to prevent its leaching. It was protected from contact with the water by a thin layer (0.2 mm) of RTV108 silicone rubber.
