Carbon nanopipettes characterize calcium release pathways in breast cancer cells

Abstract

Carbon-based nanoprobes are attractive for minimally-invasive cell interrogation but their application in cell physiology has thus far been limited. We have developed carbon nanopipettes (CNPs) with nanoscopic tips and used them to inject calcium-mobilizing messengers into cells without compromising cell viability. We identify pathways sensitive to cyclic adenosine diphosphate ribose (cADPr) and nicotinic acid adenine dinucleotide phosphate (NAADP) in breast carcinoma cells. Our findings demonstrate the superior utility of CNPs for intracellular delivery of impermeant molecules and, more generally, for cell physiology studies. The CNPs do not appear to cause any lasting damage to cells. Their advantages over the commonly used glass pipettes include smaller size, breakage and clogging resistance, and potential for multifunctionality such as concurrent injection and electrical measurements.

Effects of Deposition Conditions on the Structure and Chemical Properties of Carbon Nanopipettes

“…1d) was smooth and minimized damage to the cell membrane during intracellular injection and electrical recording. [8][9][10] In contrast, as shown in Figure 1e, the inner wall was relatively rough. As previously shown with CNTs grown in aluminum membranes, [12] this surface roughness develops when catalyst particles embed into NT walls during the carbon film formation.…”

“…CNPs have desirable mechanical and electrical properties for cell probing. They have been successfully applied for fluid injections into cells without causing cell damage or death, and for measuring the electrical response of cells to extracellular stimuli; [8][9][10] however no study of the carbon structure and its dependence on synthesis parameters has been reported. Previous studies of CNTs and nanopipes synthesized by CVD in aluminum oxide membrane templates showed that the electric conductivity, the graphitization, and the wetability of the NTs increased after annealing at high temperatures.…”

Micro-Nano Technologies for Cell Manipulation and Subcellular Monitoring

“…A better understanding of this procedure could eventually lead to commercial production and set pattern in cell handling. Scaling down further to nanofluidics has also been achieved by ingenious building of carbon nanopipettes on conventional glass pipettes (Schrlau et al, 2008). Compared to conventional glass pipettes, these structures have suggested enhanced performance for intracellular delivery and cell physiology due to their smaller size, breakage and clogging resistance.…”

Carbon-based nanoprobes for cell biology

“…calcium pathways Schrlau et al (2008b) used CNPs to inject calcium-mobilizing secondary messengers into breast cancer cells to identify intracellular calcium signaling pathways. Figure 5 shows the cytosolic calcium concentrations as functions of time in response to injections of inositol trisphosphate (IP 3 ) in intracellular solution.…”

“…5d, e, blue-green color). (Schrlau et al 2008a) In another set of experiments, Schrlau et al (2008b) demonstrated the existence of functional calcium channels sensitive to cyclic adenosine diphosphate ribose (cADPr) and nicotinic acid adenine dinucleotide phosphate (NA-ADP) in breast cancer cells. The experiments provided evidence of significant advantages of CNPs over microinjection glass pipettes.…”