More than half a century ago, Erwin Schroedinger, nobel laureate in physics, claimed that it is 'impossible to carry out experiments on single molecules or atoms'. Today, the detection, tracking and study of single molecules and atoms has become an omnipresent tool in biology, chemistry and physics alike. For example, sequencing DNA one base pair (or letter) at a time currently provides the most likely solution to fulfill the quest for a $1,000 human genome. Nevertheless, observation of a single molecule, especially with standard light microscopes requires a good deal of laboratory skills. This is mostly due to the fact that a single molecule only gives a miniscule amount of detectable signal. In fact, people using light as a probe have relied exclusively on the use of fluorescence, the emission of lower energy light following absorption of radiation at a certain energy. In this scheme, the signal from the molecule of interest can be easily separated from residual excitation light or background fluorescence simply by filtering the detected light spectrally and only detecting the color that is emitted by the molecule. In this way, it is possible to suppress unwanted signals from the billions of other molecules that are in the vicinity of the molecule of interest. As powerful as this approach has been, it also has one major limitation: it is only possible to study molecules that are highly fluorescent, i.e. emit lower energy light with high efficiency. Scientists from the ETH Zurich have recently demonstrated a major step towards the detection and study of single molecules in absorption.
Nanomaterial-based drug-delivery and nanotoxicology are two of the areas that require sophisticated methods and techniques for characterizing, testing and imaging nanoparticulate matter inside the body. Especially the potential risk factors of certain nanomaterials have become a heated topic of discussion recently. Most, if not all, toxicological studies on nanoparticles rely on current methods, practices and terminology as gained and applied in the analysis of micro- and ultrafine particles and mineral fibers. The development of novel imaging techniques that can visualize local populations of nanoparticles at nanometer resolution within the structures of cells - without destroying or damaging the cell - are therefore important. Researchers in the U.S. have now demonstrated that at ultrasonic frequencies, intracellular nanomaterial cause sufficient wave scattering that a probe outside the cell can respond to. This ultrasonic holography technique provides a non-invasive way of looking inside a cell.
'Reverse engineering' is the process of discovering the technological principles of a device or system through analysis of its structure, function and operation, often by taking it apart and analyzing its workings in detail. This approach is a common practice among industrial companies who use it to analyze the competition's products, be it cars or MP3 players, to understand where the latest product improvements come from and how individual components are made. An increasing number of scientists apply a similar approach to nature's own 'micro- and nanotechnology' systems. They believe that learning from nature's designs and engineering successes is more likely to provide the cues for designing practical nanodevices than by simply applying a 'trial and error' approach. The basic idea is that natural materials and systems can be adopted for human use beyond their original purpose in nature. Some examples of 'reverse' biophysics work and have already proven quite useful, for instance the use of individual red blood cells as reliable, ultrasensitive mechanotransducers.
With the advance of nanotechnologies the demand for ever more precise instruments that measure, map and manipulate details at the nanoscale increases as well. For instance, the study of potential distributions with nanoscale resolution becomes increasingly important. In the early days of atomic force microscopy (AFM) the scanning force microscope was used to measure charges, dielectric constants, film thickness of insulating layers, photovoltage, and electrical potential of a given surface. Then, in 1991, the concept of a scanning contact potential microscope was introduced, allowing the simultaneous measurement of topography and contact potential difference. Named the scanning surface potential microscope (SSPM) - also often referred to as Kelvin probe force microscope - this is a variation of the AFM that measures the electrostatic forces (potential) between the probe tip and the surface of a material. Compared with other AFM techniques, the lateral resolution of traditional SSPM, from submicron down to 10 nm, is much lower.
Understanding and manipulating cellular function at the level of individual molecules is within reach. One of the requirements of single molecule techniques is the ability to follow an individual molecule for sufficiently long times in solution. However, it is a challenge to cope with the effects of Brownian motion (the random motion of small particles suspended in a gas or liquid) on this time scale. To meet this challenge, more recently, biomolecules have been encapsulated inside lipid vesicles, which are themselves tethered to a surface. Now, a novel nanocontainer offers controlled permeability functionality which not only is desirable for single molecule imaging but also is a very important property for micro- and nanodevices and for delivery of drugs or imaging agents in vitro and in vivo.
Whenever you read an article about nano this or nano that, chances are you come across a large number of confusing three-letter acronyms - AFM, SFM, SEM, TEM, SPM, FIB, CNT and so on. It seems scientists earn extra kudos when they come up with a new three-letter combination. One of the most important acronyms in nanotechnology is AFM - Atomic Force Microscopy. This instrument has become the most widely used tool for imaging, measuring and manipulating matter at the nanoscale and in turn has inspired a variety of other scanning probe techniques. Originally the AFM was used to image the topography of surfaces, but by modifying the tip it is possible to measure other quantities (for example, electric and magnetic properties, chemical potentials, friction and so on), and also to perform various types of spectroscopy and analysis. Today we take a look at one of the instruments that has it all made possible. So far, over 20,000 AFM-related papers have been published; over 500 patents were issued related to various forms of scanning probe microscopes (SPM); several dozen companies are involved in manufacturing SPM and related instruments, with an annual worldwide turnover of $250-300 million, and approx. 10,000 commercial systems sold (not counting a significant number of home-built systems).
The need for 3D visualization and analysis at high spatial resolution is likely to increase as nanoscience and nanotechnology become increasingly important and nanotomography could play a key role in understanding structure, composition and physico-chemical properties at the nanoscale. Scientists from the Electron Microscopy Group at the University of Cambridge in the UK report that nanotomography is becoming an important tool in the study of the size, shape, distribution and composition of various materials, including nanomaterials.
There is a significant and growing need across the research and medical communities for low-cost, high throughput DNA separation and quantification techniques. The isolation of DNA is a prerequisite step for many molecular biology techniques and experiments. Although single molecule techniques afford extremely high sensitivity, to date, such experiments have remained within the confines of academic and research laboratories. The primary reasons for this state of affairs relate to throughput, detection efficiencies and analysis times. For example, in a conventional solution-based single molecule detection experiment, one can only detect approximately 10,000 molecules per minute, or one molecule every 6 milliseconds. While this may sound a lot, consider that a small drop of water (ca. 5 ml) contains approx. 1.67 x 10e23 molecules, that is 1.67 followed by 23 zeros. At that speed you need over 100 trillion years to detect all the water molecules in this single drop. Using a novel nanopore array developed by researchers in the UK, expect to be able to detect up to 1 million molecules simultaneously in the same 6 millisecond time window, representing an improvement in throughput of over six orders of magnitude (and bringing the timeframe for analyzing the molecules in a single water drop down to some 60 billion years - about five to six times the estimated age of the universe).