Glass micropipettes, atomic force microscope tips and nanoneedles can be used to interrogate cells, but these devices either have conical geometries that can damage cells during penetration or are incapable of continuous fluid handling. Here, we report a carbon-nanotube-based endoscope for interrogating cells, transporting fluids and performing optical and electrochemical diagnostics at the single organelle level. The endoscope, which is made by placing a multiwalled carbon nanotube (length, 50-60 µm) at the tip of a glass pipette, can probe the intracellular environment with a spatial resolution of ∼100 nm and can also access organelles without disrupting the cell. When the nanotube is filled with magnetic nanoparticles, the endoscope can be remotely manoeuvered to transport nanoparticles and attolitre volumes of fluids to and from precise locations. Because they are mounted on conventional glass micropipettes, the endoscopes readily fit standard instruments, creating a broad range of opportunities for minimally invasive intracellular probing, drug delivery and single-cell surgery.

In this review, we analyze the current hypotheses regarding energy metabolism in the neurons and astroglia. Recently, it was shown that up to 20% of the total brain's energy is provided by mitochondrial oxidation of fatty acids. However, the existing hypotheses consider glucose, or its derivative lactate, as the only main energy substrate for the brain. Astroglia metabolically supports the neurons by providing lactate as a substrate for neuronal mitochondria. In addition, a significant amount of neuromediators, glutamate and GABA, is transported into neurons and also serves as substrates for mitochondria. Thus, neuronal mitochondria may simultaneously oxidize several substrates. Astrocytes have to replenish the pool of neuromediators by synthesis de novo, which requires large amounts of energy. In this review, we made an attempt to reconcile β-oxidation of fatty acids by astrocytic mitochondria with the existing hypothesis on regulation of aerobic glycolysis. We suggest that, under condition of neuronal excitation, both metabolic pathways may exist simultaneously. We provide experimental evidence that isolated neuronal mitochondria may oxidize palmitoyl carnitine in the presence of other mitochondrial substrates. We also suggest that variations in the brain mitochondrial metabolic phenotype may be associated with different mtDNA haplogroups.

We report on a new analytical approach to intracellular chemical sensing that utilizes a surface-enhanced Raman spectroscopy (SERS)-enabled nanopipette. The probe is comprised of a glass capillary with a 100-500 nm tip coated with gold nanoparticles. The fixed geometry of the gold nanoparticles allows us to overcome the limitations of the traditional approach for intracellular SERS using metal colloids. We demonstrate that the SERS-enabled nanopipettes can be used for in situ analysis of living cell function in real time. In addition, SERS functionality of these probes allows tracking of their localization in a cell. The developed probes can also be applied for highly sensitive chemical analysis of nanoliter volumes of chemicals in a variety of environmental and analytical applications.

Nanoscale multifunctional carbon probes facilitate cellular studies due to their small size, which makes it possible to interrogate organelles within living cells in a minimally invasive fashion. However, connecting nanotubes to macroscopic devices and constructing an integrated system for the purpose of fluid and electrical signal transfer is challenging, as is often the case with nanoscale components. We describe a non-catalytic chemical vapor deposition based method for batch fabrication of integrated multifunctional carbon nanopipettes (CNPs) with tip diameters much smaller (10-30 nm) than previously reported (200 nm and above) and approaching those observed for multiwalled carbon nanotubes. This eliminates the need for complicated attachment/assembly of nanotubes into nanofluidic devices. Variable tip geometries and structures were obtained by controlled deposition of carbon inside and outside quartz pipettes. We have shown that the capillary length and gas flow rate have a marked effect on the carbon deposition. This gives us a flexible protocol, useful for growing carbon layers of different thicknesses at selective locations on a glass pipette to yield a large variety of cellular probes in bulk quantities. The CNPs possess an open channel for fluid transfer with the carbon deposited inside at 875 degrees C behaving like an amorphous semiconductor. Vacuum annealing of the CNP tips at temperatures up to 2000 degrees C yields graphitic carbon structures with an increase in conductivity of two orders of magnitude. Penetration of the integrated carbon nanoprobes into cells was shown to produce minimal Ca(2+) signals, fast recovery of basal Ca(2+) levels and no adverse activation of the cellular metabolism during interrogation times as long as 0.5-1 h.

Surface‐enhanced Raman spectroscopy (SERS) is a promising and powerful label free technique for high resolution analysis of single cells. For intracellular analysis, there is a need for SERS‐active nanoprobes that are minimally invasive to cells, do not affect cell viability, and provide reproducible signals. This work reviews the state‐of‐the‐art tools currently available for intracellular SERS. Various types of SERS probes are considered, including colloidal gold and silver nanoparticles, metallized optical fibers, and tip‐enhanced Raman probes. We also discuss recently developed SERS‐active nanopipettes implemented on the basis of pulled glass microcapillaries. Finally, the critical aspects of selecting an optimal SERS nanoprobe for single‐cell analysis depending on a particular application are summarized. Copyright © 2012 John Wiley & Sons, Ltd.