Open AccessA novel Ca2+ indicator for long-term tracking of intracellular calcium flux
Author(s) -
Jinfang Liao,
Deven Patel,
Qin Zhao,
Ruogu Peng,
Haitao Guo,
Zhenjun Diwu
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0161
Subject(s) - chemistry , biophysics , calcium , fluorescence , stimulation , intracellular , incubation , biochemistry , biology , optics , physics , organic chemistry , neuroscience
The major drawback of using Fluo-4 AM is that it requires an organic anion transporter inhibitor, such as probenecid, to prevent leakage. This can hinder the studies that require extended monitoring time and longer cellular retention. To address the issue, a novel Ca 2+ indicator, Calbryte 520 AM, was developed. We compared the performance of Fluo-4 AM and Calbryte 520 AM following prolonged incubation periods after cell loading. Cells loaded with Calbryte 520 AM retained the dye for up to 24 h while exhibiting significant fluorescence brightness and superior F max /F 0 ratios (F max : fluorescence intensity upon stimulation; F 0 : intensity before stimulation). It demonstrated that the longer retention of Calbryte 520 AM can be exploited to accommodate for the extended time required when monitoring calcium dynamics.