There is, I think, something ethereal about i —the square root of minus one. I remember first hearing about it at school. It seemed an odd beast at that time—an intruder hovering on the edge of reality.

Usually familiarity dulls this sense of the bizarre, but in the case of i it was the reverse: over the years the sense of its surreal nature intensified. It seemed that it was impossible to write mathematics that described the real world in …

I and i

Molecular excitation by the simultaneous absorption of two photons provides intrinsic three-dimensional resolution in laser scanning fluorescence microscopy. The excitation of fluorophores having single-photon absorption in the ultraviolet with a stream of strongly focused subpicosecond pulses of red laser light has made possible fluorescence images of living cells and other microscopic objects. The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation. This technique also provides unprecedented capabilities for three-dimensional, spatially resolved photochemistry, particularly photolytic release of caged effector molecules.

https://materias.df.uba.ar/TemasNano2013/files/2013/07/1990_Denk_Science_Two-photon-fluo-microscopy.pdf

Two-Photon Laser Scanning Fluorescence Microscopy

Research on fluorescent semiconductor nanocrystals (also known as quantum dots or qdots) has evolved over the past two decades from electronic materials science to biological applications. We review current approaches to the synthesis, solubilization, and functionalization of qdots and their applications to cell and animal biology. Recent examples of their experimental use include the observation of diffusion of individual glycine receptors in living neurons and the identification of lymph nodes in live animals by near-infrared emission during surgery. The new generations of qdots have farreaching potential for the study of intracellular processes at the single-molecule level, high-resolution cellular imaging, long-term in vivo observation of cell trafficking, tumor targeting, and diagnostics.

/pdf/quantum-dots-for-live-cells-in-vivo-imaging-and-diagnostics-3y9ayecoh8.pdf

Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics

The interest in nanoscale materials stems from the fact that new properties are acquired at this length scale and, equally important, that these properties * To whom correspondence should be addressed. Phone, 404-8940292; fax, 404-894-0294; e-mail, mostafa.el-sayed@ chemistry.gatech.edu. † Case Western Reserve UniversitysMillis 2258. ‡ Phone, 216-368-5918; fax, 216-368-3006; e-mail, burda@case.edu. § Georgia Institute of Technology. 1025 Chem. Rev. 2005, 105, 1025−1102

Chemistry and properties of nanocrystals of different shapes.

One of the fastest moving and most exciting interfaces of nanotechnology is the use of quantum dots (QDs) in biology. The unique optical properties of QDs make them appealing as in vivo and in vitro fluorophores in a variety of biological investigations, in which traditional fluorescent labels based on organic molecules fall short of providing long-term stability and simultaneous detection of multiple signals. The ability to make QDs water soluble and target them to specific biomolecules has led to promising applications in cellular labelling, deep-tissue imaging, assay labelling and as efficient fluorescence resonance energy transfer donors. Despite recent progress, much work still needs to be done to achieve reproducible and robust surface functionalization and develop flexible bioconjugation techniques. In this review, we look at current methods for preparing QD bioconjugates as well as presenting an overview of applications. The potential of QDs in biology has just begun to be realized and new avenues will arise as our ability to manipulate these materials improves.

Quantum dot bioconjugates for imaging, labelling and sensing

The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have two-photon action cross sections as high as 47,000 Goeppert-Mayer units, by far the largest of any label used in multiphoton microscopy. We visualized quantum dots dynamically through the skin of living mice, in capillaries hundreds of micrometers deep. We found no evidence of blinking (fluorescence intermittency) in solution on nanosecond to millisecond time scales.

https://science.sciencemag.org/content/sci/300/5624/1434.full.pdf

Water-Soluble Quantum Dots for Multiphoton Fluorescence Imaging in Vivo

Two-photon fluorescence microscopy (2PM)1 enables scientists in various fields including neuroscience2,3, embryology4, and oncology5 to visualize in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of 2PM within the mouse brain to the cortical layer, and imaging subcortical structures currently requires the removal of overlying brain tissue3 or the insertion of optical probes6,7. Here we demonstrate non-invasive, high resolution, in vivo imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy (3PM) at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein (RFP)-labeled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher order nonlinear excitation overcomes the limitations of 2PM, enabling biological investigations to take place at greater depth within tissue.

/pdf/in-vivo-three-photon-microscopy-of-subcortical-structures-4m5ry8wq1o.pdf

In vivo three-photon microscopy of subcortical structures within an intact mouse brain

Nanocrystals or quantum dots of the IV−VI semiconductors PbS, PbSe, and PbTe provide unique properties for investigating the effects of strong confinement on electrons and phonons. The degree of confinement of charge carriers can be many times stronger than in most II−VI and III−V semiconductors, and lead salt nanostructures may be the only materials in which the electronic energies are determined primarily by quantum confinement. This Account briefly reviews recent research on lead salt quantum dots.

Lead Salt Quantum Dots: the Limit of Strong Quantum Confinement

Self-similar propagation of ultrashort, parabolic pulses in a laser resonator is observed theoretically and experimentally. This constitutes a new type of pulse shaping in mode-locked lasers: in contrast to the well-known static (solitonlike) and breathing (dispersion-managed soliton) pulse evolutions, asymptotic solutions to the nonlinear wave equation that governs pulse propagation in most of the laser cavity are observed. Stable self-similar pulses exist with energies much greater than can be tolerated in solitonlike pulse shaping, and this has implications for practical lasers.

https://arxiv.org/pdf/physics/0402013.pdf

Self-similar evolution of parabolic pulses in a laser.

In the course of the past several years, a new level of understanding has been achieved about conditions for the existence, stability, and generation of spatiotemporal optical solitons, which are nondiffracting and nondispersing wavepackets propagating in nonlinear optical media. Experimentally, effectively two-dimensional (2D) spatiotemporal solitons that overcome diffraction in one transverse spatial dimension have been created in quadratic nonlinear media. With regard to the theory, fundamentally new features of light pulses that self-trap in one or two transverse spatial dimensions and do not spread out in time, when propagating in various optical media, were thoroughly investigated in models with various nonlinearities. Stable vorticity-carrying spatiotemporal solitons have been predicted too, in media with competing nonlinearities (quadratic–cubic or cubic–quintic). This article offers an up-to-date survey of experimental and theoretical results in this field. Both achievements and outstanding difficulties are reviewed, and open problems are highlighted. Also briefly described are recent predictions for stable 2D and 3D solitons in Bose–Einstein condensates supported by full or low-dimensional optical lattices.

https://iopscience.iop.org/article/10.1088/1464-4266/7/5/R02/pdf

Frank W. Wise

Papers

Water-Soluble Quantum Dots for Multiphoton Fluorescence Imaging in Vivo

In vivo three-photon microscopy of subcortical structures within an intact mouse brain

Lead Salt Quantum Dots: the Limit of Strong Quantum Confinement

Self-similar evolution of parabolic pulses in a laser.

Spatiotemporal optical solitons