ArticleCold Spring Harbor protocols2012
Construction of flow chambers for polarized total internal reflection fluorescence microscopy (polTIRFM) motility assays.
Article in Cold Spring Harbor protocols, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
4 citing papers in PubMed, 3 citations in OpenAlex.
- Angular measurements of the dynein ring reveal a stepping mechanism dependent on a flexible stalk.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- Preparation of segmented microtubules to study motions driven by the disassembling microtubule ends.Journal of visualized experiments : JoVE · 2014Article
- The polarized total internal reflection fluorescence microscopy (polTIRFM) twirling filament assay.Cold Spring Harbor protocols · 2012Article
- The polarized total internal reflection fluorescence microscopy (polTIRFM) processive motility assay for myosin V.Cold Spring Harbor protocols · 2012Article
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Authors and funding
5 authors.
Funding
Abstract
Polarized total internal reflection fluorescence microscopy (polTIRFM) can be used to detect the spatial orientation and rotational dynamics of single molecules. polTIRFM determines the three-dimensional angular orientation and the extent of wobble of a fluorescent probe bound to the macromolecule of interest. This protocol describes how to construct sample chambers (flow chambers) for polTIRFM motility assays. Each chamber can hold ∼20 µL of solution. To flow a solution through the chamber, the solution is added to the chamber with a pipette while wicking out the previous contents with filter paper. Each end of the coverslip should extend beyond the edge of the slide to support the pipette tip and filter paper. The flow rate can be roughly controlled by adjusting the contact area between the filter paper and the solution. The chambers can be used for investigating the motility of myosin V in vitro with the processive motility assay, as well as for assessing the motility of actin using the twirling assay.
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