RRC ID 72622
Author Hideyuki Yaginuma, Yasushi Okada
Title Live cell imaging of metabolic heterogeneity by quantitative fluorescent ATP indicator protein, QUEEN-37C
Abstract Adenosine triphosphate (ATP) is often referred as the energy currency of the cell. Yet, non-invasive, real-time, and quantitative measurement of its concentration in living mammalian cells has been difficult. Here we report an improved fluorescent ATP indicator protein, QUEEN-37C, which is optimized for measuring ATP concentration in living mammalian cells. Absolute value of the ATP concentration can be estimated from the ratiometric fluorescence imaging, and its accuracy was verified by the luciferase assay. Since QUEEN-37C enables the single-cell measurement of ATP concentration, we can not only measure its mean but its distribution in the cell population, which revealed that the ATP concentration is tightly regulated in most cells. We also noted the positive correlations in the ATP concentration among adjacent cells in epithelial cell sheet and mouse embryonic stem cell colonies. Thus, QUEEN-37C would serve as a new tool for the investigation of the single cell heterogeneity of metabolic states.
DOI 10.1101/2021.10.08.463131
Resource
DNA material pN1/G3BP1-iRFP (RDB17742) pLBS.CAG (RDB17751) pRSET-QUE2m (RDB17754) pRS-QUE7mu (RDB17770) pT2MP.EF-pHlu (RDB17787) pRSAN-QUE37C (RDB17788) pRSAN-QUE20C (RDB17791) pRSET-QUE7mu R122A (RDB17793) pLBS.EF NLS-pHlu (RDB17809) pLBS.EF-iRFP (RDB17822) pT2MP.EF-QUE37C (RDB17825) pLBS.EF/miRFP670-hGem110Nt (RDB17826) pLBS.EF/mScarlet(i)-hCdt100Nt (RDB17828) pLBS.EF (RDB17829) pLBS-mScarlet (RDB17830) pLBS-EF-QUE37C (RDB17831) QUE37C/pLBS.CAG (RDB17832) pRSET-QUEEN 7mu A81D (RDB17834) pRSET-QUEEN 7mu A81I (RDB17835) pRSET-QUEEN 7mu E83A (RDB17836) pRSET-QUEEN 7mu D89A (RDB17837) pRSET-QUEEN 7mu V90D (RDB17838) pRSET-QUEEN 7mu A81E (RDB17839) pRSET-QUEEN 7mu I88D (RDB17840) pRSET QUEEN7mu K26A (RDB17841) pRSET QUEEN7mu K28A (RDB17842) pRSET QUEEN7mu R51A (RDB17843) pRSET QUEEN7mu K53A (RDB17844) pRSET QUEEN7mu K54A (RDB17845) pRSET QUEEN7mu K57A (RDB17846) pRSET QUEEN7mu K71A (RDB17847) pRSET QUEEN7mu K74A (RDB17848) pRSET QUEEN7mu R84A (RDB17849) pRSET QUEEN7mu R92A (RDB17850) pRSET QUEEN7mu K94A (RDB17851) pRSET QUEEN7mu K97A (RDB17852) pRSET QUEEN7mu R99A (RDB17853) pRSET QUEEN7mu R102A (RDB17854) pRSET QUEEN7mu R103A (RDB17855) pRSET QUEEN7mu K114A (RDB17856) pRSET QUEEN7mu R115A (RDB17857) pRSET QUEEN7mu K121A (RDB17858) pRSET-QUEEN 7mu R126A (RDB17859) pRSET-QUEEN 7mu R126I (RDB17860) pRS-QUEEN 84BP (RDB17861) pRSAN-QUEEN BPEK (RDB17862) pRSAN-QUEEN BPEE (RDB17863) pRSAN-QUEEN BPKK (RDB17864) pRSAN-QUEEN BPRR (RDB17865)