Super-Resolved Traction Force Microscopy (STFM)

Colin‐York, Huw; Shrestha, Dilip; Felce, James H.; Waithe, Dominic; Moeendarbary, Emad; Davis, Simon J.; Eggeling, Christian; Fritzsche, Marco

doi:10.1021/acs.nanolett.6b00273

Cited by 100 publications

(127 citation statements)

References 22 publications

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“…Related to this, the molecular spatial composition of cell adhesions were also mapped using iPALM 153 . Further, new applications of STED to traction force microscopy allow for increased resolution for measuring the forces that cells exert on extracellular substrates 154 .…”

Section: Super-resolution Investigations Of Biological Membranesmentioning

confidence: 99%

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

2017

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Lipids and the membranes they form are fundamental building blocks of cellular life, and their geometry and chemical properties distinguish membranes from other cellular environments. Collective processes occurring within membranes strongly impact cellular behavior and biochemistry, and understanding these processes presents unique challenges due to the often complex and myriad interactions between membrane components. Super-resolution microscopy offers a significant gain in resolution over traditional optical microscopy, enabling the localization of individual molecules even in densely labeled samples and in cellular and tissue environments. These microscopy techniques have been used to examine the organization and dynamics of plasma membrane components, providing insight into the fundamental interactions that determine membrane functions. Here, we broadly introduce the structure and organization of the mammalian plasma membrane and review recent applications of super-resolution microscopy to the study of membranes. We then highlight some inherent challenges faced when using super-resolution microscopy to study membranes, and we discuss recent technical advancements that promise further improvements to super-resolution microscopy and its application to the plasma membrane.

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Section: Super-resolution Investigations Of Biological Membranesmentioning

confidence: 99%

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

2017

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“…Localization microscopy must ensure that no two molecules emit inside the diffraction spot size, what can be achieved by, e.g., turning on and off the molecules emission. Indeed, using super‐resolution techniques, traction force microscopy spatial resolution and accuracy of force reconstruction have been significantly improved . In our case, each nanowire is more than a diffraction spot size apart, so we do not encounter this problem.…”

Section: Optical Measurementsmentioning

confidence: 91%

“…Over the years, many techniques have been applied or specifically developed for measuring cell adhesion forces . Among them, flow chamber experiments have been widely used but they mostly capture shear forces in fluids.…”

Section: Introductionmentioning

confidence: 99%

Nanowire Arrays as Force Sensors with Super‐Resolved Localization Position Detection: Application to Optical Measurement of Bacterial Adhesion Forces

et al. 2018

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The design and application of indium phosphide (InP) nanowire arrays to acquire Xylella fastidiosa bacterial cell vector force maps are discussed. The nanowire deflections are measured with subdiffraction localization confocal laser scanning microscopy (CLSM). The nanowire mechanical stability in air and liquid media as well as methods to average out thermally induced oscillations are investigated. The accuracy of center determination of the CLSM reflected laser intensity profile at nanowire apex is studied using Gaussian fitting and localization microscopy techniques. These results show that the method is reliable for measuring nanowire displacements above ≈25 nm. Corresponding force ranges probed by this method can be customized depending on nanowire geometry and array configuration. The method is applied to explore X. fastidiosa cell adhesion forces on the InP nanowire surface, and in situ probes the effect of N‐acetylcysteine on adhered cells. Future perspectives for application of this method in microbiology studies are also outlined.

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“…This resolution is achieved in all three dimensions using a cross‐correlation analysis of the intensity profile in the

x y

‐plane and by using the diffraction pattern calibration along the z ‐axis (Gosse & Croquette, ). For optical imaging, choices of fiducial markers include fluorescent dyes (Vaughan et al ., ), fluorescent beads (Polte et al ., ; Georgieva et al ., ; Icha et al ., ; Tafteh et al ., ), nonfluorescent beads (Steffen et al ., ; Nugent‐Glandorf & Perkins, ; Capitanio et al ., ; Vaughan et al ., ), microfabricated patterns (Carter et al ., ; Sabass et al ., ; Lee et al ., ; Polio et al ., ; Colin‐York et al ., ), quantum dots (Betzig et al ., ; Kukulski et al ., ) and gold nanoparticles (Betzig et al., ; York et al ., ; Kukulski et al., ; Bon et al ., ). Multimodal imaging requires fiducial markers that can be tracked with each imaging method.…”

Section: Introductionmentioning

confidence: 99%

Estimation of microscope drift using fluorescent nanodiamonds as fiducial markers

Colomb

Czerski

et al. 2017

Journal of Microscopy

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Fiducial markers are used to correct the microscope drift and should be photostable, be usable at multiple wavelengths and be compatible for multimodal imaging. Fiducial markers such as beads, gold nanoparticles, microfabricated patterns and organic fluorophores lack one or more of these criteria. Moreover, the localization accuracy and drift correction can be degraded by other fluorophores, instrument noise and artefacts due to image processing and tracking algorithms. Estimating mechanical drift by assuming Gaussian distributed noise is not suitable under these circumstances. Here we present a method that uses fluorescent nanodiamonds as fiducial markers and uses an improved maximum likelihood algorithm to estimate the drift with both accuracy and precision within the range 1.55-5.75 nm.

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Super-Resolved Traction Force Microscopy (STFM)

Cited by 100 publications

References 22 publications

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

Super-Resolution Microscopy: Shedding Light on the Cellular Plasma Membrane

Nanowire Arrays as Force Sensors with Super‐Resolved Localization Position Detection: Application to Optical Measurement of Bacterial Adhesion Forces

Estimation of microscope drift using fluorescent nanodiamonds as fiducial markers

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