Evidence map›Paper›PMID 35604180›Full record

ArticleJournal of visualized experiments : JoVE2022

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins.

Yazgan Tuna, Amer Al-Hiyasat, Jonathon Howard

Open access · greenAbstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
0.8field-weighted citation impact, top 34% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

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.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 8 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

3 authors at 1 institution in 2 countries.

Yazgan TunaDepartment of Molecular Biophysics & Biochemistry, Yale University.
Amer Al-HiyasatDepartment of Molecular Biophysics & Biochemistry, Yale University.
Jonathon HowardDepartment of Molecular Biophysics & Biochemistry, Yale University; Department of Physics, Yale University; jonathon.howard@yale.edu.
Institute of Molecular Biology and Biophysics · RU

Funding

Dendrite structure: Data-Driven Models to Bridge from Molecules to MorphologyR01NS118884 · NINDS · YALE UNIVERSITY · PI HOWARD, JONATHON · 2021 to 2025
$2.0M
Control of microtubule length by polymerases and depolymerasesR01GM110386 · NIGMS · YALE UNIVERSITY · PI HOWARD, JONATHON · 2014 to 2017
$1.6M
Microtubule Severing and Regrowth by SpastinR01GM139337 · NIGMS · YALE UNIVERSITY · PI HOWARD, JONATHON · 2020 to 2023
$1.3M
NIGMS NIH HHS R01 GM110386NIGMS NIH HHS R01 GM139337NINDS NIH HHS R01 NS118884
6 · The paper itself

Abstract

Several techniques have been employed for the direct visualization of cytoskeletal filaments and their associated proteins. Total-internal-reflection-fluorescence (TIRF) microscopy has a high signal-to-background ratio, but it suffers from photobleaching and photodamage of the fluorescent proteins. Label-free techniques such as interference reflection microscopy (IRM) and interferometric scattering microscopy (iSCAT) circumvent the problem of photobleaching but cannot readily visualize single molecules. This paper presents a protocol for combining IRM with a commercial TIRF microscope for the simultaneous imaging of microtubule-associated proteins (MAPs) and dynamic microtubules in vitro. This protocol allows for high-speed observation of MAPs interacting with dynamic microtubules. This improves on existing two-color TIRF setups by eliminating both the need for microtubule labeling and the need for several additional optical components, such as a second excitation laser. Both channels are imaged on the same camera chip to avoid image registration and frame synchronization problems. This setup is demonstrated by visualizing single kinesin molecules walking on dynamic microtubules.

Indexed as

Microtubule-Associated ProteinsMicrotubulesMicroscopy, FluorescenceMicroscopy, InterferencePhotobleachingMicrotubule-Associated Proteins

Identifiers

PMID35604180
PMCPMC12212060
OpenAlexW4225402042

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.