Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane

Taylor, Richard W.; Mahmoodabadi, Reza Gholami; Rauschenberger, Verena; Gießl, Andreas; Schambony, Alexandra; Sandoghdar, Vahid

doi:10.1101/401133

Cited by 11 publications

(13 citation statements)

References 47 publications

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“…Phase imaging [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] provides morphological phasecontrast of transparent samples and is widely used in various fields, especially in biological science, because morphological features of micrometre-scale specimens provide valuable information on complex biological phenomena. Quantitative phase imaging (QPI) [1][2][3][4][5][6] is the most powerful method for studying cellular morphology among various phase imaging methods, such as phase-contrast 7 and differential-interference-contrast 8 imaging, because it is able to accurately measure the optical phase delay (OPD) caused by a sample.…”

Section: Introductionmentioning

confidence: 99%

“…In addition, wide-field scattering imaging techniques using dark-field (DF) 14,15 or interferometric scattering (iSCAT) 16,17 have also used the concept of differential image analysis to observe dynamically changing small signals originating from fast-moving nanoscale scattering objects in a slowly moving microscale environment. They have been mostly used for investigating simple biomimicking systems 16,17 and, more recently, applied to measure gold nanoparticles on cell membranes 18,19 . However, the limited dynamic range of DF imaging causes decreased sensitivity in the presence of large-OPD objects (>1 rad) such as cells.…”

Section: Introductionmentioning

confidence: 99%

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Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

2021

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Quantitative phase imaging (QPI) with its high-contrast images of optical phase delay (OPD) maps is often used for label-free single-cell analysis. Contrary to other imaging methods, sensitivity improvement has not been intensively explored because conventional QPI is sensitive enough to observe the surface roughness of a substrate that restricts the minimum measurable OPD. However, emerging QPI techniques that utilize, for example, differential image analysis of consecutive temporal frames, such as mid-infrared photothermal QPI, mitigate the minimum OPD limit by decoupling the static OPD contribution and allow measurement of much smaller OPDs. Here, we propose and demonstrate supersensitive QPI with an expanded dynamic range. It is enabled by adaptive dynamic range shift through a combination of wavefront shaping and dark-field QPI techniques. As a proof-of-concept demonstration, we show dynamic range expansion (sensitivity improvement) of QPI by a factor of 6.6 and its utility in improving the sensitivity of mid-infrared photothermal QPI. This technique can also be applied for wide-field scattering imaging of dynamically changing nanoscale objects inside and outside a biological cell without losing global cellular morphological image information.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

2021

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“…Tracking of free particles variable in size quickly passing in the field of view is then almost impossible. A significant amelioration of ring-shaped PSFs (12,13) was recently proposed (21), allowing nanoparticle tracking in living cells, but is limited in axial range (a few hundreds of nanometers) and precision (5 to 6 nm).…”

Section: Introductionmentioning

confidence: 99%

Parallel, linear, and subnanometric 3D tracking of microparticles with Stereo Darkfield Interferometry

et al. 2021

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While crucial for force spectroscopists and microbiologists, three-dimensional (3D) particle tracking suffers from either poor precision, complex calibration, or the need of expensive hardware, preventing its massive adoption. We introduce a new technique, based on a simple piece of cardboard inserted in the objective focal plane, that enables simple 3D tracking of dilute microparticles while offering subnanometer frame-to-frame precision in all directions. Its linearity alleviates calibration procedures, while the interferometric pattern enhances precision. We illustrate its utility in single-molecule force spectroscopy and single-algae motility analysis. As with any technique based on back focal plane engineering, it may be directly embedded in a commercial objective, providing a means to convert any preexisting optical setup in a 3D tracking system. Thanks to its precision, its simplicity, and its versatility, we envision that the technique has the potential to enhance the spreading of high-precision and high-throughput 3D tracking.

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“…Given recent advances in camera technology this will likely not pose a limitation for much longer. For example, interferometric scattering (iSCAT) microscopy, which relies on collecting scattered light from the sample rather than fluorescence emission, allows frame rates of multiple kilo Hertz covering most of the range of dynamic processes in biology ( 97 – 99 ).…”

Section: Fluorescence Fluctuation Based Approaches To Assess Simultanmentioning

confidence: 99%

Quantitative Bio-Imaging Tools to Dissect the Interplay of Membrane and Cytoskeletal Actin Dynamics in Immune Cells

2021

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Cellular function is reliant on the dynamic interplay between the plasma membrane and the actin cytoskeleton. This critical relationship is of particular importance in immune cells, where both the cytoskeleton and the plasma membrane work in concert to organize and potentiate immune signaling events. Despite their importance, there remains a critical gap in understanding how these respective dynamics are coupled, and how this coupling in turn may influence immune cell function from the bottom up. In this review, we highlight recent optical technologies that could provide strategies to investigate the simultaneous dynamics of both the cytoskeleton and membrane as well as their interplay, focusing on current and future applications in immune cells. We provide a guide of the spatio-temporal scale of each technique as well as highlighting novel probes and labels that have the potential to provide insights into membrane and cytoskeletal dynamics. The quantitative biophysical tools presented here provide a new and exciting route to uncover the relationship between plasma membrane and cytoskeletal dynamics that underlies immune cell function.

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Interferometric scattering microscopy reveals microsecond nanoscopic protein motion on a live cell membrane

Cited by 11 publications

References 47 publications

Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

Adaptive dynamic range shift (ADRIFT) quantitative phase imaging

Parallel, linear, and subnanometric 3D tracking of microparticles with Stereo Darkfield Interferometry

Quantitative Bio-Imaging Tools to Dissect the Interplay of Membrane and Cytoskeletal Actin Dynamics in Immune Cells

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