Fluorescent Keima pH Titration

Cells overexpressing fluorescent protein keima were incubated in a series of buffers with pH values ranging from 4.12 to 7.97, then the fluorescent signal of keima was measured under the conditions of excitation of 440 and 550 nm and emission of 610 nm using Nikon ECLIPSE Ti-U epi-fluorescence microscope. Then the fluorescent intensity of keima was measured by NIS-Elements F 3.0 software^{37 (link)}. The pH titration curves and fitting equation were obtained by the negative correlation between the pH value and the fluorescent intensity ratio of 550 nm/440 nm of keima protein.

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Su Y., Ren H., Tang M., Zheng Y., Zhang B., Wang C., Hou X., Niu Z., Wang Z., Gao X., Gao L., Jiang H., Chen Z., Luo T, & Sun Q. (2021). Role and dynamics of vacuolar pH during cell-in-cell mediated death. Cell Death & Disease, 12(1), 119.

Publication 2021

ECLIPSE Ti-U epi-fluorescence microscope

Buffers Cells F elements Fluorescence microscope Protein Titration

Corresponding Organization :

Other organizations : University of Science and Technology of China, Beijing Shijitan Hospital, Capital Medical University, Beijing Jiaotong University, Chinese Academy of Medical Sciences & Peking Union Medical College

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Variable analysis

independent variables

PH values ranging from 4.12 to 7.97

dependent variables

Fluorescent signal of keima measured under the conditions of excitation of 440 and 550 nm and emission of 610 nm

control variables

Cells overexpressing fluorescent protein keima
Nikon ECLIPSE Ti-U epi-fluorescence microscope
NIS-Elements F 3.0 software

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