title: Evaluating nanoparticle sensor design for intracellular pH measurements.
abstract: Particle-based nanosensors have over the past decade been designed for optical fluorescent-based ratiometric measurements of pH in living cells. However, quantitative and time-resolved intracellular measurements of pH in endosomes and lysosomes using particle nanosensors are challenging, and there is a need to improve measurement methodology. In the present paper, we have successfully carried out time-resolved pH measurements in endosomes and lyosomes in living cells using nanoparticle sensors and show the importance of sensor choice for successful quantification. We have studied two nanoparticle-based sensor systems that are internalized by endocytosis and elucidated important factors in nanosensor design that should be considered in future development of new sensors. From our experiments it is clear that it is highly important to use sensors that have a broad measurement range, as erroneous quantification of pH is an unfortunate result when measuring pH too close to the limit of the sensitive range of the sensors. Triple-labeled nanosensors with a pH measurement range of 3.2-7.0, which was synthesized by adding two pH-sensitive fluorophores with different pK(a) to each sensor, seem to be a solution to some of the earlier problems found when measuring pH in the endosome-lysosome pathway.
authors.affiliation: DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of Denmark, Building 423, 2800 Lyngby, Denmark.
authors.forename: Rikke V
authors.initials: RV
authors.lastname: Benjaminsen
authors.forename: Honghao
authors.initials: H
authors.lastname: Sun
authors.forename: Jonas R
authors.initials: JR
authors.lastname: Henriksen
authors.forename: Nynne M
authors.initials: NM
authors.lastname: Christensen
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Almdal
authors.forename: Thomas L
authors.initials: TL
authors.lastname: Andresen
mesh_terms: D002138:Calibration; D002470:Cell Survival; D011992:Endosomes / Q000737:chemistry*; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D013816:Thermodynamics; D013997:Time Factors
chemical_list: D011108:Polymers