Biosensors (Basel), 2021, 11(9)

FLIM-Based Intracellular and Extracellular pH Measurements Using Genetically Encoded pH Sensor

The determination of pH in live cells and tissues is of high importance in physiology and cell biology. In this report, we outline the process of the creation of SypHerExtra, a genetically encoded fluorescent sensor that is capable of measuring extracellular media pH in a mildly alkaline range. SypHerExtra is a protein created by fusing the previously described pH sensor SypHer3s with the neurexin transmembrane domain that targets its expression to the cytoplasmic membrane. We showed that with excitation at 445 nm, the fluorescence lifetime of both SypHer3s and SypHerExtra strongly depend on pH. Using FLIM microscopy in live eukaryotic cells, we demonstrated that SypHerExtra can be successfully used to determine extracellular pH, while SypHer3s can be applied to measure intracellular pH. Thus, these two sensors are suitable for quantitative measurements using the FLIM method, to determine intracellular and extracellular pH in a range from pH 7.5 to 9.5 in different biological systems.

Goryashchenko AS, Pakhomov AA, Ryabova AV, Romanishkin ID, Maksimov EG, Orsa AN, Serova OV, Mozhaev AA, Maksimova MA, Martynov VI, Petrenko AG, Deyev IE

IBCH: 9453
Ссылка на статью в журнале: https://www.mdpi.com/2079-6374/11/9/340
Кол-во цитирований на 06.2024: 12
Данные статьи проверены модераторами 2021-09-27

Список научных проектов, где отмечена публикация

  1. Physiological role of gene insrr in pancreatic beta-cells (January 7, 2019 — December 31, 2021). Deyev S.M.. Grant, RFBR.
  2. - (January 6, 2020 — December 31, 2022). Goryashchenko A.S., Deyev I.E.. Grant, RFBR.
  3. 19-73-20194 (January 6, 1970). . Сommercial.