The delivery of PEBBLE nanosensors to measure the intracellular environment

Abstract

Cellular introduction of PEBBLEs (photonic explorers for bioanalysis with biologically localized embedding) has been investigated by a wide variety of methods in a range of cell types. These methods include surface functionalization with CPPs (cell-penetrating peptides), pinocytosis, commercial lipid transfection agents, cytochalasin D, picoinjection, and Gene gun bombardment. This paper will overview several of the most popular methods used for the introduction of PEBBLE nanosensors to the cellular environment and discuss the efficacy of the techniques.

The development and in vitro characterisation of an intracellular nanosensor responsive to reactive oxygen species

“…Although phagocytosis has been demonstrated as a successful means of delivering the nanosensors into cells this limits the application of the ROS nanosensor only to those cells capable of phagocytosis. One of the areas of future work will be to deliver the nanosensors into other cell types via alternative delivery methods such as those described in the literature (Webster et al, 2007). Monitoring ROS generation over extended periods of time (one of the key advantages the PEBBLE technology offers over conventional cell loading with fluorescent dyes) may enable greater resolution of the temporal and spatial dynamics of ROS in various biomedical scenarios.…”

“…phagocytosis (Webster et al, 2007). In this study ROS sensitive PEBBLEs were delivered into macrophage cells through phagocytosis.…”

Sensing Inside the Living Cells

“…Much like with fl uorescent organic dyes, there is no general solution of the problem of delivery of nanoparticles into the cells. Currently used methods include picoinjections, conjugation with cationic peptides, liposomal delivery, sequestration into macrophages and gene gun (Webster et al 2007 ). Each of these methods can be tried but no one of them is perfect or generally applicable.…”

Electrochemical and optical sensing of reactive oxygen species: pathway to an integrated intracellular and extracellular measurement platform

“…PEBBLE nanosensors for calcium [29], zinc [30], glucose [31] and pH [32] have all been reported. Numerous methods for introducing them into the intracellular environment have been reported including linking nanosensors to the cell penetrating TAT peptide [29], physical insertion via a gene gun [33] and utilizing innate cellular phagocytosis [34]. All of these methods have their limitations [33].…”

“…Numerous methods for introducing them into the intracellular environment have been reported including linking nanosensors to the cell penetrating TAT peptide [29], physical insertion via a gene gun [33] and utilizing innate cellular phagocytosis [34]. All of these methods have their limitations [33]. However, a recent innovation that delivered nanosensors via lipid transfection [33] has been shown to offer a reliable method that was suitable for both immortalized cell lines and primary cells.…”