Nanometer‐scale measurements using video light microscopy

Schnapp, Bruce J.; Gelles, Jeff; Sheetz, Michael P.

doi:10.1002/cm.970100109

Cited by 69 publications

(51 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Reconstituted type I collagen fibrils are well resolved by DIC that has been used to study cell/matrix interaction in collagen gels (Petroll & Ma 2003). In addition, video-enhanced DIC has been used to estimate the diameter of biological filaments and structures well below the optical diffraction limit Schnapp et al 1988). Thus, we surmised that the loss of monomer from degrading collagen fibrils would likely be reflected in the DIC signal, even when the fibril diameters become very small.…”

Section: (B) Enzyme and Substratementioning

confidence: 99%

Mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase: implications for load-bearing matrix growth and stability

Bhole

Flynn

Liles

et al. 2009

Phil. Trans. R. Soc. A.

119

109

View full text Add to dashboard Cite

There has been great interest in understanding the methods by which collagen-based load-bearing tissue is constructed, grown and maintained in vertebrate animals. To date, the responsibility for this process has largely been placed with mesenchymal fibroblastic cells that are thought to fully control the morphology of load-bearing extracellular matrix (ECM). However, given clear limitations in the ability of fibroblastic cells to precisely place or remove single collagen molecules to sculpt tissue, we have hypothesized that the material itself must play a critical role in the determination of the form of structural ECM. We here demonstrate directly, using live, dynamic, differential interference contrast imaging, that mechanically strained networks of collagen fibrils, exposed to collagenase (Clostridium histolyticum), degrade preferentially. Specifically, unstrained fibrils are removed 'quickly', while strained fibrils persist significantly longer. The demonstration supports the idea that collagen networks are mechanosensitive in that they are stabilized by mechanical strain. Thus, collagen molecules (together with their complement enzymes) may comprise the basis of a smart, load-adaptive, structural material system. This concept has the potential to drastically simplify the assumed role of the fibroblast, which would need only to provide ECM molecules and mechanical force to sculpt collagenous tissue.

show abstract

Section: (B) Enzyme and Substratementioning

confidence: 99%

Mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase: implications for load-bearing matrix growth and stability

Bhole

Flynn

Liles

et al. 2009

Phil. Trans. R. Soc. A.

119

109

View full text Add to dashboard Cite

show abstract

“…The general idea of single-particle tracking (SPT) is to determine the position of an individual particle by fitting its diffraction-limited image on a CCD camera to the known point-spread function of the microscope or, more pragmatically, by fitting it to a twodimensional Gaussian. Associating the center of the fit with the position of the particle allows determination of the spatial position with an accuracy far beyond the classical diffraction limit of light microscopy and to follow the diffusion of this particle with high precision [1,3,[16][17][18][19][20][21]. Fascinating experiments have been reported using SPT in combination with fluorescently labeled particles.…”

Section: Introductionmentioning

confidence: 99%

Setup for single-particle orbit tracking: artifacts and corrections

2012

View full text Add to dashboard Cite

We report on an experimental setup for single-particle orbit tracking, which allows following fluorescent nanoparticles for more than 10 min with a temporal resolution of 4 ms and a dynamic position accuracy of better than 10 nm. On a model sample--20 nm sized fluorescent polymer beads in glycerol--we will illustrate how artifacts caused by unavoidable experimental shortcomings (might) obscure the experimental result and how misinterpretations can be prevented.

show abstract

“…(73) The technique was further improved in later years. (74,75) A large body of work on imaging and tracking of single lipid molecules on artificial and cell membranes was further catalyzed by Schutz and coworkers. (76) Later on, the technique was further developed by to track the particles 3-D with different strategies.…”

Section: Single-particle Trackingmentioning

confidence: 99%

Membrane Dynamics: Fluorescence Spectroscopy

Subburaj

Gallego

García‐Sáez

2000

Encyclopedia of Analytical Chemistry

View full text Add to dashboard Cite

Membranes are dynamic lipophilic structures that constitute a selective barrier essential for the smooth functioning of cellular machinery. Their complex and highly heterogeneous composition, as well as the intricate diffusion patterns that they display, have been intensely studied in the past decades. Several innovative microscopy techniques have recently been developed to gain insight into the basis of membrane dynamics, interactions, and molecular organization. In this article, we introduce four pioneer fluorescence‐based approaches that have substantially broadened our overall understanding of membranes: fluorescence correlation spectroscopy (FCS), single particle tracking (SPT), fluorescence recovery after photobleaching (FRAP), and 6‐lauroyl‐2‐(dimethylamino)‐naphtalene (LAURDAN) microscopy. We discuss each technique in detail, explaining their principles, methodology, and applications in the research of the dynamic processes controlled by membranes.

show abstract

Nanometer‐scale measurements using video light microscopy

Cited by 69 publications

References 15 publications

Mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase: implications for load-bearing matrix growth and stability

Mechanical strain enhances survivability of collagen micronetworks in the presence of collagenase: implications for load-bearing matrix growth and stability

Setup for single-particle orbit tracking: artifacts and corrections

Membrane Dynamics: Fluorescence Spectroscopy

Contact Info

Product

Resources

About