An Optimized MicroPET Imaging Method for the Distribution and Synergies of Natural Products

Image-Based Marker-Free Screening of GABAA Agonists, Antagonists, and Modulators

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/2472555219887142 ◽

2019 ◽

Vol 25 (5) ◽

pp. 458-470 ◽

Cited By ~ 1

Author(s):

Benjamin Rappaz ◽

Pascal Jourdain ◽

Damiano Banfi ◽

Fabien Kuttler ◽

Pierre Marquet ◽

...

Keyword(s):

Natural Products ◽

Fluorescent Dyes ◽

Methodological Approach ◽

Free Cell ◽

Digital Holographic Microscopy ◽

Imaging Method ◽

Chemical Structures ◽

Agonist Action ◽

Electrophysiological Measurements ◽

Marker Free

The ionotropic GABAA receptors represent the main target for different groups of widely used drugs having hypnotic and anxiolytic effects. So far, most approaches used to assess GABA activity involve invasive low -throughput electrophysiological techniques or rely on fluorescent dyes, preventing the ability to conduct noninvasive and thus nonperturbing screens. To address this limitation, we have developed an automated marker-free cell imaging method, based on digital holographic microscopy (DHM). This technology allows the automatically screening of compounds in multiple plates without having to label the cells or use special plates. This methodological approach was first validated by screening the GABAA receptor expressed in HEK cells using a selection of active compounds in agonist, antagonist, and modulator modes. Then, in a second blind screen of a library of 3041 compounds (mostly composed of natural products), 5 compounds having a specific agonist action on the GABAA receptor were identified. The hits validated from this unbiased screen were the natural products muscimol, neurosteroid alphaxalone, and three compounds belonging to the avermectin family, all known for having an agonistic effect on the GABAA receptor. The results obtained were exempt from false negatives (structurally similar unassigned hits), and false-positive hits were detected and discarded without the need for performing electrophysiological measurements. The outcome of the screen demonstrates the applicability of our screening by imaging method for the discovery of new chemical structures, particularly regarding chemicals interacting with the ionotropic GABAA receptor and more generally with any ligand-gated ion channels and transporters.

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Weak Beam Imaging and Diffraction Studies of Stacking Faults in Silicon

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100092918 ◽

1976 ◽

Vol 34 ◽

pp. 590-591

Author(s):

T. Y. Tan ◽

W. K. Tice

Keyword(s):

Fast Neutron ◽

Rapid Determination ◽

Stacking Faults ◽

Imaging Method ◽

Large Reduction ◽

Dislocation Loops ◽

Fringe Spacing ◽

Weak Beam ◽

Kinematical Theory

In studying ion implanted semiconductors and fast neutron irradiated metals, the need for characterizing small dislocation loops having diameters of a few hundred angstrom units usually arises. The weak beam imaging method is a powerful technique for analyzing these loops. Because of the large reduction in stacking fault (SF) fringe spacing at large sg, this method allows for a rapid determination of whether the loop is faulted, and, hence, whether it is a perfect or a Frank partial loop. This method was first used by Bicknell to image small faulted loops in boron implanted silicon. He explained the fringe spacing by kinematical theory, i.e., ≃l/(Sg) in the fault fringe in depth oscillation. The fault image contrast formation mechanism is, however, really more complicated.

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Performance and applications of the holography electron microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100151222 ◽

1993 ◽

Vol 51 ◽

pp. 1078-1079

Author(s):

Akira Tonomura

Keyword(s):

Electron Beam ◽

Electron Microscope ◽

Phase Measurement ◽

Electron Holography ◽

Imaging Method ◽

High Contrast ◽

Magnetic Lens ◽

High Brightness ◽

Holographic Imaging ◽

Dynamic Observation

Electron holography is a two-step imaging method. However, the ultimate performance of holographic imaging is mainly determined by the brightness of the electron beam used in the hologram-formation process. In our 350kV holography electron microscope (see Fig. 1), the decrease in the inherently high brightness of field-emitted electrons is minimized by superposing a magnetic lens in the gun, for a resulting value of 2 × 109 A/cm2 sr. This high brightness has lead to the following distinguished features. The minimum spacing (d) of carrier fringes is d = 0.09 Å, thus allowing a reconstructed image with a resolution, at least in principle, as high as 3d=0.3 Å. The precision in phase measurement can be as high as 2π/100, since the position of fringes can be known precisely from a high-contrast hologram formed under highly collimated illumination. Dynamic observation becomes possible because the current density is high.

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