microscopy (3)

Drops in Cells...

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Liquidated3672 (2021), Theodore Lee Jones, CallMeTed.com.

Topics: Applied Physics, Biology, Microscopy, Molecules

A major challenge in cell biology remains to unravel is how cells control their biochemical reaction cycles. For instance, how do they regulate gene expression in response to stress? How does their metabolism change when resources are scarce? Control theory has proven useful in understanding how networks of chemical reactions can robustly tackle those and other tasks.1 The essential ingredients in such approaches are chemical feedback loops that create control mechanisms similar to the circuits that regulate, for example, the temperature of a heating system, the humidity of an archive, or the pH of a fermentation tank.

Theories for the control of biochemical reactions have largely focused on homogeneous, well-stirred environments. However, macromolecules inside cells are often highly organized in space by specialized subunits called organelles. Some organelles, such as the cell nucleus, are bound by a membrane. By contrast, another class of organelles—biomolecular condensates—show the hallmark physical properties of liquid-like droplets, and they provide chemically distinct environments for biochemical reactions.2–4

Such droplets can act as microreactors for biochemical reactions in a living cell (see figure 1). Their liquid nature sustains the fast diffusion of reactants while their specific composition gives rise to the partitioning of reactants in or out of the droplets. In general, the concentrations of reactants inside condensates differ from the concentrations outside. Those differences modify reaction fluxes, which, in turn, can dramatically affect reaction yield and other properties of chemical reactions. Just how such modified fluxes govern the biochemistry inside cells remains poorly understood.

Drops in Cells, Christoph Weber, Christoph Zechner, Physics Today

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NEMS Photothermal Microscopy...

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Topics: Microscopy, Nanotechnology, NEMS, Physics, Research

Single-molecule microscopy has become an indispensable tool for biochemical analysis. The capability of characterizing distinct properties of individual molecules without averaging has provided us with a different perspective for the existing scientific issues and phenomena. Recently, super-resolution fluorescence microscopy techniques have overcome the optical diffraction limit by the localization of molecule positions. However, the labeling process can potentially modify the intermolecular dynamics. Based on the highly sensitive nanomechanical photothermal microscopy reported previously, we propose optimizations on this label-free microscopy technique toward localization microscopy. A localization precision of 3 Å is achieved with gold nanoparticles, and the detection of polarization-dependent absorption is demonstrated, which opens the door for further improvement with polarization modulation imaging.

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FIG. 2. (a) Schematic of the measurement setup. BE: beam expander. M: mirror. WP: waveplate. LP: linear polarizer. BS: beam splitter. PD: photodetector/power meter. DM: dichroic mirror. ID: iris diaphragm. CCD: charge-coupled device camera. APD: avalanche photodiode detector. (b) The transduction scheme of the trampoline resonator. (c) SEM image of the trampoline resonator.

J. Appl. Phys. 128, 134501 (2020); https://doi.org/10.1063/5.0014905

Nanoelectromechanical photothermal polarization microscopy with 3 Å localization precision, Miao-Hsuan Chien and Silvan Schmid, Journal of Applied Physics

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Holographic Microscopy...

Figure

FIG. 1. (a) Schematic of an inline digital holographic microscope. In a typical setup, a collimated laser (light red) illuminates a sample, which scatters light (dark red wavefronts). The transmitted and scattered light passes through an objective and tube lens, which focuses the light onto a digital camera. (b) A hologram of a polystyrene particle obtained from an inline holographic microscope.

Topics: Holography, Optical Physics, Microscopy, Modern Physics

In the past few years, the venerable field of holographic microscopy has been revitalized by computational data analysis. It is now possible to fit a generative (forward) model of scattering directly to experimentally obtained holograms of complex microscopic objects. This approach enables precision measurements: it allows the motion of colloidal particles and biological organisms to be tracked with nanometer-scale precision and their optical properties to be inferred particle by particle. In this Perspective, we discuss how the model-based inference approach to holographic microscopy has opened up new applications. We also discuss how it must evolve to meet the needs of emerging applications that demand lower systematic uncertainties and higher precision. In this context, we present some new results on how modeling the optical train of the microscope can enable better measurements of the positions of spherical and nonspherical colloidal particles. Finally, we discuss how machine learning might play a role in future advances. Though we do not exhaustively catalog all the developments in this field, we show a few examples and some new results that spotlight open questions and opportunities.

 

Precise measurements in digital holographic microscopy by modeling the optical train, Ronald Alexander, Brian Leahy, Vinothan N. Manoharan, Journal of Applied Physics

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