Non Animal Testing Database

Two-photon ratiometric probe for real-time intracellular pH monitoring

November 2021
University of Sydney, Sydney, Australia
Designing a two-photon ratiometric pH-sensitive nanoprobe for real-time monitoring of intracellular pH in a biological environment is of great importance for a better understanding of the pathogenesis of diseases and the design of intracellular drug delivery-based systems. However, the development of such a probe remains a challenge. Here, the design and development of the two-photon ratiometric carbon dot-based nanoprobe are shown for the first time and its capability to monitor and quantify the intracellular cytoplasmic pH value, in real-time is demonstrated. The nanoprobe is comprised of a fluorescent carbon dot functionalized with a pH-responsive fluorescein dye (FACD). With increasing pH, FACD exhibits a clear ratiometric change in the emission intensity ratio, with sensitivity across a wide pH range in both extracellular and intracellular compartments. FACD is non-toxic to human adipose stem cells in cell imaging experimental conditions and exhibits remarkable thermal stability, photostability, and two-photon near-infrared excitation capability. Using real-time dual-channel two-photon confocal microscopy this study demonstrates the great potential of FACD as an efficient nanoprobe with high accuracy for the intracellular sensing of pH in living adipose stem cells seeded on either cell-culture dishes or on a 3D printed bioactive scaffold.
Two-photon ratiometric carbon dot-based probe for real-time intracellular pH monitoring in 3D environment
Pooria Lesani, Hala Zreiqat
Added on: 12-15-2022
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