Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy.

Nanosecond temporal resolution enables new methods for wide-field imaging like time-of-flight, gated detection, and fluorescence lifetime. The optical efficiency of existing approaches, however, presents challenges for low-light applications common to fluorescence microscopy and single-molecule imaging. We demonstrate the use of Pockels cells for wide-field image gating with nanosecond temporal resolution and high photon collection efficiency. Two temporal frames are obtained by combining a Pockels cell with a pair of polarizing beam-splitters. We show multi-label fluorescence lifetime imaging microscopy (FLIM), single-molecule lifetime spectroscopy, and fast single-frame FLIM at the camera frame rate with 103-105 times higher throughput than single photon counting. Finally, we demonstrate a space-to-time image multiplexer using a re-imaging optical cavity with a tilted mirror to extend the Pockels cell technique to multiple temporal frames. These methods enable nanosecond imaging with standard optical systems and sensors, opening a new temporal dimension for wide-field low-light microscopy.

Design for a 10 keV multi-pass transmission electron microscope.

Multi-pass transmission electron microscopy (MPTEM) has been proposed as a way to reduce damage to radiation-sensitive materials. For the field of cryo-electron microscopy (cryo-EM), this would significantly reduce the number of projections needed to create a 3D model and would allow the imaging of lower-contrast, more heterogeneous samples. We have designed a 10 keV proof-of-concept MPTEM. The column features fast-switching gated electron mirrors which cause each electron to interrogate the sample multiple times. A linear approximation for the multi-pass contrast transfer function (CTF) is developed to explain how the resolution depends on the number of passes through the sample.

Multi-pass transmission electron microscopy.

Feynman once asked physicists to build better electron microscopes to be able to watch biology at work. While electron microscopes can now provide atomic resolution, electron beam induced specimen damage precludes high resolution imaging of sensitive materials, such as single proteins or polymers. Here, we use simulations to show that an electron microscope based on a multi-pass measurement protocol enables imaging of single proteins, without averaging structures over multiple images. While we demonstrate the method for particular imaging targets, the approach is broadly applicable and is expected to improve resolution and sensitivity for a range of electron microscopy imaging modalities, including, for example, scanning and spectroscopic techniques. The approach implements a quantum mechanically optimal strategy which under idealized conditions can be considered interaction-free.

Iterative creation and sensing of twisted light.

The iterative interaction of a photon with a sample can lead to increased sensitivity in measuring the properties of the samples, such as its refractive index or birefringence. Here we show that this principle can also be used to generate and sense states of light. In particular, we demonstrate a technique to generate states with high orbital angular momentum using a single-vortex phase plate (VPP). This is accomplished by placing the phase plate in a self-imaging cavity such that light interacts with it multiple times; for an ideal phase plate, this is equivalent to iterative applications of the angular momentum operator. Using a discrete VPP, we show that our setup realizes a high-dimensional generalization of the Pauli matrix σx, and that the created states show sub-diffraction limited features that might find applications in structured illumination microscopy.

Multi-pass microscopy.

Microscopy of biological specimens often requires low light levels to avoid damage. This yields images impaired by shot noise. An improved measurement accuracy at the Heisenberg limit can be achieved exploiting quantum correlations. If sample damage is the limiting resource, an equivalent limit can be reached by passing photons through a specimen multiple times sequentially. Here we use self-imaging cavities and employ a temporal post-selection scheme to present full-field multi-pass polarization and transmission micrographs with variance reductions of 4.4±0.8 dB (11.6±0.8 dB in a lossless setup) and 4.8±0.8 dB, respectively, compared with the single-pass shot-noise limit. If the accuracy is limited by the number of detected probe particles, our measurements show a variance reduction of 25.9±0.9 dB. The contrast enhancement capabilities in imaging and in diffraction studies are demonstrated with nanostructured samples and with embryonic kidney 293T cells. This approach to Heisenberg-limited microscopy does not rely on quantum state engineering.

An atomically thin matter-wave beamsplitter.

Matter-wave interferometry has become an essential tool in studies on the foundations of quantum physics and for precision measurements. Mechanical gratings have played an important role as coherent beamsplitters for atoms, molecules and clusters, because the basic diffraction mechanism is the same for all particles. However, polarizable objects may experience van der Waals shifts when they pass the grating walls, and the undesired dephasing may prevent interferometry with massive objects. Here, we explore how to minimize this perturbation by reducing the thickness of the diffraction mask to its ultimate physical limit, that is, the thickness of a single atom. We have fabricated diffraction masks in single-layer and bilayer graphene as well as in a 1 nm thin carbonaceous biphenyl membrane. We identify conditions to transform an array of single-layer graphene nanoribbons into a grating of carbon nanoscrolls. We show that all these ultrathin nanomasks can be used for high-contrast quantum diffraction of massive molecules. They can be seen as a nanomechanical answer to the question debated by Bohr and Einstein of whether a softly suspended double slit would destroy quantum interference. In agreement with Bohr's reasoning we show that quantum coherence prevails, even in the limit of atomically thin gratings.

Ultrafast oscilloscope based on laser-triggered field emitters.

Laser-triggered electron emission from sharp metal tips has been demonstrated in recent years as a high brightness, ultrafast electron source. Its possible applications range from ultrafast electron microscopy to laser-based particle accelerators to electron interferometry. The ultrafast nature of the emission process allows for the sampling of an instantaneous radio frequency (RF) voltage that has been applied to a field emitter. For proof-of-concept, we use an RF signal derived from our laser's repetition rate, mapping a 9.28 GHz signal in 22.4 fs steps with 28 mv accuracy.

Ultrafast Time-Resolved Photoelectric Emission.

The emission times of laser-triggered electrons from a sharp tungsten tip are directly characterized under ultrafast, near-infrared laser excitation at Keldysh parameters of 6.6<γ<19.1. Emission delays up to 10 fs are observed, which are inferred from the energy gain of photoelectrons emitted into a synchronously driven microwave cavity. Few femtosecond timing resolution is achieved in a configuration capable of measuring timing shifts up to 55 ps. The technique can also be used to measure the microwave phase inside the cavity with a precision below 70 fs upon the energy resolved detection of a single electron.

Experimental methods of molecular matter-wave optics.

We describe the state of the art in preparing, manipulating and detecting coherent molecular matter. We focus on experimental methods for handling the quantum motion of compound systems from diatomic molecules to clusters or biomolecules.Molecular quantum optics offers many challenges and innovative prospects: already the combination of two atoms into one molecule takes several well-established methods from atomic physics, such as for instance laser cooling, to their limits. The enormous internal complexity that arises when hundreds or thousands of atoms are bound in a single organic molecule, cluster or nanocrystal provides a richness that can only be tackled by combining methods from atomic physics, chemistry, cluster physics, nanotechnology and the life sciences.We review various molecular beam sources and their suitability for matter-wave experiments. We discuss numerous molecular detection schemes and give an overview over diffraction and interference experiments that have already been performed with molecules or clusters.Applications of de Broglie studies with composite systems range from fundamental tests of physics up to quantum-enhanced metrology in physical chemistry, biophysics and the surface sciences.Nanoparticle quantum optics is a growing field, which will intrigue researchers still for many years to come. This review can, therefore, only be a snapshot of a very dynamical process.

Real-time single-molecule imaging of quantum interference.

The observation of interference patterns in double-slit experiments with massive particles is generally regarded as the ultimate demonstration of the quantum nature of these objects. Such matter-wave interference has been observed for electrons, neutrons, atoms and molecules and, in contrast to classical physics, quantum interference can be observed when single particles arrive at the detector one by one. The build-up of such patterns in experiments with electrons has been described as the "most beautiful experiment in physics". Here, we show how a combination of nanofabrication and nano-imaging allows us to record the full two-dimensional build-up of quantum interference patterns in real time for phthalocyanine molecules and for derivatives of phthalocyanine molecules, which have masses of 514 AMU and 1,298 AMU respectively. A laser-controlled micro-evaporation source was used to produce a beam of molecules with the required intensity and coherence, and the gratings were machined in 10-nm-thick silicon nitride membranes to reduce the effect of van der Waals forces. Wide-field fluorescence microscopy detected the position of each molecule with an accuracy of 10 nm and revealed the build-up of a deterministic ensemble interference pattern from single molecules that arrived stochastically at the detector. In addition to providing this particularly clear demonstration of wave-particle duality, our approach could also be used to study larger molecules and explore the boundary between quantum and classical physics.

Immobilization of zinc porphyrin complexes on pyridine-functionalized glass surfaces.

In order to immobilize sublimable and fluorescent dye molecules on transparent surfaces for the detection of far field molecular interference experiments, we investigate the potential of pyridine-functionalized glass substrates as coordination sites for the zinc complex of tetraphenylporphyrin (ZnTPP). Borosilicate glass is functionalized with 4-(6-(ethoxydimethylsilyl)hexyloxy)pyridine in order to cover the glass surface with pyridine subunits. ZnTPP molecules are deposited by sublimation through mechanical masks of various sizes in a high-vacuum chamber. The resulting micropatterns are analyzed using epifluorescence microscopy which also allows us to define a measure for the quality of molecular immobilization. We observe a reduced mobility and an increased efficiency for the trapping of ZnTPP on pyridine-functionalized surfaces.

Quantum physics meets biology.

Quantum physics and biology have long been regarded as unrelated disciplines, describing nature at the inanimate microlevel on the one hand and living species on the other hand. Over the past decades the life sciences have succeeded in providing ever more and refined explanations of macroscopic phenomena that were based on an improved understanding of molecular structures and mechanisms. Simultaneously, quantum physics, originally rooted in a world-view of quantum coherences, entanglement, and other nonclassical effects, has been heading toward systems of increasing complexity. The present perspective article shall serve as a "pedestrian guide" to the growing interconnections between the two fields. We recapitulate the generic and sometimes unintuitive characteristics of quantum physics and point to a number of applications in the life sciences. We discuss our criteria for a future "quantum biology," its current status, recent experimental progress, and also the restrictions that nature imposes on bold extrapolations of quantum theory to macroscopic phenomena.

Wave and particle in molecular interference lithography.

The wave-particle duality of massive objects is a cornerstone of quantum physics and a key property of many modern tools such as electron microscopy, neutron diffraction or atom interferometry. Here we report on the first experimental demonstration of quantum interference lithography with complex molecules. Molecular matter-wave interference patterns are deposited onto a reconstructed Si(111) 7x7 surface and imaged using scanning tunneling microscopy. Thereby both the particle and the quantum wave character of the molecules can be visualized in one and the same image. This new approach to nanolithography therefore also represents a sensitive new detection scheme for quantum interference experiments.