In the early 1870s, the German physicist Ernst Karl Abbé formulated a rigorous criterion for being able to resolve two objects in a light microscope. According to his equation, the best resolution achievable with visible light is about 200 nanometers. This theoretical resolution limit of conventional optical imaging methodology was the primary factor motivating the development of recent higher-resolution scanning probe techniques. The interaction of light with an object results in the generation of what is called 'near-field' and 'far-field' light components. The far-field light propagates through space in an unconfined manner and is the visible light utilized in conventional light microscopy. The near-field (or evanescent) light consists of a nonpropagating field that exists near the surface of an object at distances less than a single wavelength of light. So called near-field microscopy beats light's diffraction limit by moving the source very close to the subject to be imaged. When the first theoretical work on a new technique called "scanning near-field optical microscopy" (SNOM or NSOM) appeared in the 1980's, Abbé's classical diffraction limit was overcome, and resolution even down to single molecule level became feasible. However, light microscopy is still the only way to observe the interior of whole, or even living, cells. The use of fluorescent dyes makes it possible to selectively obtain images of individual cell components, for example, proteins. Today, the wavelength dogma has been overcome with the development of the stimulated emission depletion (STED) microscope. Now, the German team that developed STED is reporting layer-by-layer light microscopic nanoscale images of cells and without having to prepare thin sections with a technique called optical 3D far-field microscopy. They use a chemical marker for fluorescence nanoscopy that relies on single-molecule photoswitching.
"Children begin to learn by seeing, hearing, tasting and, above all, by touching. In a very similar approach, we are currently learning to orient ourselves in the nanoworld by 'feeling' materials - not with our fingers, but with microscopes that allow us to probe these materials with atomic resolution." (Robert W. Stark, LMU Munich in "Getting a feeling for the nanoworld"). Researchers' ability to engineer materials and achieve superior electronic, thermal, magnetic, and mechanical properties depends on tools that can identify and characterize material components and their spatial arrangement at the nanoscale. Equally important, understanding structure and function relationships in biological systems also demands tools that can probe structural properties with molecular resolution. Atomic force microscopes (AFM) are the most widely used tools to image matter at the nanoscale. Due to its mechanical operation, the AFM can in principle also perform nanomechanical measurements. This aspect of the AFM has been explored by researchers over the past two decades. However, current state-of-the-art techniques are very slow (it takes about one second for the AFM tip to approach, push into and retract from the surface of a material) and they apply rather large forces during the measurement process that damage the tip and the sample. Researchers at Harvard and Stanford universities have developed a specially designed AFM cantilever tip, the torsional harmonic cantilever (THC), which offers orders of magnitude improvements in temporal resolution, spatial resolution, indentation and mechanical loading compared to conventional tools. With high operating speed, increased force sensitivity and excellent lateral resolution, this tool facilitates practical mapping of nanomechanical properties.
You might remember our Spotlight from a few months ago (25 years of scanning probe microscopy and no standards yet) where we gave an overview of how scanning probe microscopy has flourished over the past 25 years. The most versatile implementation of the scanned probe principle is the atomic force microscope (AFM). It has become one of the foremost tools for imaging, measuring and manipulating matter at the nanoscale. The essential part of an AFM is a microscale cantilever with a sharp tip (probe) at its end that is used to scan the specimen surface. The cantilever is typically silicon or silicon nitride with a tip radius of curvature on the order of nanometers. When the tip is brought into proximity of a sample surface, forces between the tip and the sample lead to a deflection of the cantilever according to Hooke's law. A multi-segment photodiode measures the deflection via a laser beam, which is reflected on the cantilever surface. Because there are so many promising areas in nanotechnology and biophysics which can be examined by AFM (force spectroscopy on DNA, muscle protein titin, polymers or more complex structures like bacteria flagella, 3-D imaging, etc. ) the availability of instruments is crucial, especially for new groups and young scientists with limited funds. The price tag of AFMs runs in the hundreds of thousand s of dollars, though. Until now, AFM heads are made of metal materials by conventional milling, which restricts the design and increases the costs. German researchers have shown that rapid prototyping can be a quicker and less costly alternative to conventional manufacturing.
Cells are the smallest 'brick' in life's building structures. Every living organism is made of cells. Individual cells carry their own DNA and have their own life cycle. Considering that larger organisms, such as humans, are basically huge, organized cell cooperatives, the study of individual live cells is a hugely important scientific task. Among the most significant technical challenges for performing successful live-cell imaging experiments is to maintain the cells in a healthy state and functioning normally on the microscope stage while being illuminated. Especially if scientists want to look into cellular processes that occur in cells in their natural state and that cannot be observed by traditional cytological methods. It is well known that cells move, grow, duplicate, and move from point A to point B. Up to now people studied these mechanical properties with optical microscopes because it is the most common and simple method, very efficient, a very well developed and advanced technology. However, with optical microscopes detection is limited to objects no smaller than the wavelengths of the visible region of light, roughly between 400 and 700 nanometers. Distances or movement smaller than this range cannot be seen with these instruments. Researchers in Kyoto, Japan have applied a near-field optical approach to measure cell mechanics and were able to show intriguing data of nanoscale cell membrane dynamics associated with different phenomena of the cell's life, such as cell cycle and cell death.
It has been 25 years since the scanning tunneling microscope (STM) was invented, followed four years later by the atomic force microscope, and that's when nanoscience and nanotechnology really started to take off. Various forms of scanning probe microscopes based on these discoveries are essential for many areas of today's research. Scanning probe techniques have become the workhorse of nanoscience and nanotechnology research. Given the 25-year development timeframe, it is surprising that even today there is no generally accepted standard for scanning probe microscopy (SPM). There is no unified SPM terminology, nor is there a standard for data management and treatment, making access and processing of SPM data collected by different types of instruments an error-prone exercise. SPM standardization has only recently begun as part of an effort by the International Organization for Standardization (ISO) the largest developer of industrial standards. Meanwhile the development of SPM instruments and analysis software continues, increasing the already large family of scanning probe microscopy.
Magnetic resonance imaging (MRI) is a powerful imaging technology that serves as a non-invasive method to render images of the inside of an object. It is primarily used in medical imaging to demonstrate pathological or other physiological alterations of living tissues. MRI also has uses outside of the medical field, for instance as a non-destructive testing method to characterize the quality of products such as produce and timber. Conventional MRI usually operates at the scale of millimeters to micrometers - 3 micrometers at best - which is good enough for the mostly medical diagnostic purposes it is used for. Researchers have now shown that the imaging of nuclear spins using magnetic resonance, the basis for MRI, can be pushed to sub-100nm resolution into the nanoscale realm. They demonstrated that using an emerging technique based on force detection, they can image nuclear spins with a sensitivity that is 60,000 times better than MRI. The resolution is about 30 times better than the most advanced conventional MRI imaging. By improving this technique, researchers will be able to push deeper into the nanometer regime and approach the capability needed for direct three-dimensional imaging of individual macromolecules.
It its more than 25 years of existence, Scanning Tunneling Microscopy (STM) has predominantly brought us extremely detailed images of matter at the molecular and atomic level. STM - not to be confused with the scanning electron microscope (SEM) - is a non-optical microscope that scans an electrical probe over a surface to be imaged to detect a weak electric current flowing between the tip and the surface. The STM allows scientists to visualize regions of high electron density and hence infer the position of individual atoms and molecules on the surface of a lattice. Researchers have now taken a further step by using STM to perform real-time single-molecule imaging of an entire chemical reaction. Many chemical reactions are catalyzed by metal complexes, and insight into their mechanisms is essential for the design of future catalysts. These new findings have demonstrated that the STM approach to studying chemical reactions in a dynamic environment can provide valuable information about reaction mechanisms and rates, as well as catalyst activity and stability.
Synthesis of carbon nanotubes (CNTs) is a rapidly advancing field, but there is a lot that researchers don't know about how nanotubes form and grow. Synthesis, while rapidly developing, is currently the weakest link for most nanotube applications, with high yield and high precision diameter and chirality control being important goals. Historically, in situ characterization tools have accelerated progress in synthesis for many advanced materials, and there is widespread recognition that in situ tools have the potential to improve CNT synthesis as well. Ideally one would like to detect individual nanotubes and ensembles as they grow and measure their physical properties while imposing minimal constraints on the synthesis method. In other words, with a good understanding of the synthesis process we would be better able to control the product. It is feasible that by actually observing nanotubes as they grow one will gain a better understanding of the growth process and also better characterize the grown product. Greater control over the physical characteristics of the nanotube product is essential to enable many applications, as well as many fundamental studies. Although chemical vapor deposition (CVD) is now a very standard method to synthesize CNTs, there aren't really standard in situ tools to characterize nanotubes during growth. Researchers in Canada have now shown how global Raman imaging (GRI) can be used to characterize the CVD growth of CNTs in situ and in real time.