Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH

Zhang, Liqiang1; Su, Fengyu1; Kong, Xiangxing1; Lee, Fred1; Day, Kevin1; Gao, Weimin1; Vecera, Mary E.1; Sohr, Jeremy M.1; Buizer, Sean1; Tian, Yanqing1,2; Meldrum, Deirdre R.1

2016

10.1039/c6ra06468j

RSC Advances   影响因子和分区

2046-2069

Extracellular pH has a strong effect on cell metabolism and growth. Precisely detecting extracellular pH with high throughput is critical for cell metabolism research and fermentation applications. In this research, a series of ratiometric fluorescent pH sensitive polymers are developed and the ps-pH-neutral is characterized as the best one for exclusive detection of extracellular pH. Poly(N-(2-hydroxypropyl) methacrylamide) (PHPMA) is used as the host polymer to increase the water solubility of the pH sensitive polymer without introducing cell toxicity. The fluorescent emission spectra from the polymeric sensor under excitation at the isosbestic point 455 nm possess two fluorescence peaks at 475 nm and 505 nm, which have different responding trends to pH. This enables the polymer to detect pH using fluorescent maxima at 475 nm and 505 nm (I-475 (nm)/I-505 (nm)) ratiometrically. The cell impermeability ensures the sensor can solely detect the environmental pH. The sensor is tested to detect the extracellular pH of bacteria or eukaryotic cells in high throughput assays using a microplate reader. Results showed that the pH sensor can be used for high throughput detection of extracellular pH with high repeatability and low photobleaching effect.

SCI ; EI

英语

其他

NIH Common Fund LINCS program[5 U01 CA164250]

Chemistry

Chemistry, Multidisciplinary

WOS:000377254800011

ROYAL SOC CHEMISTRY

20162202443044

Cells ; Cytology ; Emission spectroscopy ; Fluorescence ; Metabolism ; pH sensors ; Photobleaching ; Physiology ; Polymers ; Throughput

Biological Materials and Tissue Engineering:461.2 ; Biology:461.9 ; Light/Optics:741.1 ; Chemistry, General:801.1 ; Polymeric Materials:815.1

Web of Science

1.Arizona State Univ, Biodesign Inst, Ctr Biosignatures Discovery Automat, 1001 S McAlister Ave,POB 876501, Tempe, AZ 85287 USA
2.South Univ Sci & Technol China, Dept Mat Sci & Engn, 1088 Xueyuan Rd, Shenzhen 518055, Guangdong, Peoples R China

Zhang, Liqiang,Su, Fengyu,Kong, Xiangxing,et al. Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH[J]. RSC Advances,2016,6(52):46134-46142.

Zhang, Liqiang.,Su, Fengyu.,Kong, Xiangxing.,Lee, Fred.,Day, Kevin.,...&Meldrum, Deirdre R..(2016).Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH.RSC Advances,6(52),46134-46142.

Zhang, Liqiang,et al."Ratiometric fluorescent pH-sensitive polymers for high-throughput monitoring of extracellular pH".RSC Advances 6.52(2016):46134-46142.