A live cell imaging-based assay for tracking particle uptake by clathrin-mediated endocytosis.

Grant Ashby, Kayla E Keng, Carl C Hayden, Jeanne C Stachowiak

Abstract read

In one paragraph

Article in Methods in enzymology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it

0cells of the map it votes in

1citing papers in PubMed

–field-weighted citation impact

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

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3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

Poly-ubiquitylated transmembrane proteins outcompete other cargo for limited space inside clathrin-coated vesicles.Cell reports · 2025
Article

4 · The record

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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

4 authors.

Grant AshbyDepartment of Biomedical Engineering, The University of Texas at Austin.

Kayla E KengDepartment of Biomedical Engineering, The University of Texas at Austin.

Carl C HaydenDepartment of Biomedical Engineering, The University of Texas at Austin.

Jeanne C StachowiakDepartment of Biomedical Engineering, The University of Texas at Austin; Department of Chemical Engineering, The University of Texas at Austin. Electronic address: jcstach@austin.utexas.edu.

Funding

Protein Networks as Synergistic Drivers of Membrane RemodelingR35GM139531 · NIGMS · UNIVERSITY OF TEXAS AT AUSTIN · PI Jeanne Casstevens Stachowiak · 2021 to 2026

$4.0M

NIGMS NIH HHS R35 GM139531

6 · The paper itself

Abstract

A popular strategy for therapeutic delivery to cells and tissues is to encapsulate therapeutics inside particles that cells internalize via endocytosis. The efficacy of particle uptake by endocytosis is often studied in bulk using flow cytometry and Western blot analysis and confirmed using confocal microscopy. However, these techniques do not reveal the detailed dynamics of particle internalization and how the inherent heterogeneity of many types of particles may impact their endocytic uptake. Toward addressing these gaps, here we present a live-cell imaging-based method that utilizes total internal reflection fluorescence microscopy to track the uptake of a large ensemble of individual particles in parallel, as they interact with the cellular endocytic machinery. To analyze the resulting data, we employ an open-source tracking algorithm in combination with custom data filters. This analysis reveals the dynamic interactions between particles and endocytic structures, which determine the probability of particle uptake. In particular, our approach can be used to examine how variations in the physical properties of particles (size, targeting, rigidity), as well as heterogeneity within the particle population, impact endocytic uptake. These data impact the design of particles toward more selective and efficient delivery of therapeutics to cells.

Indexed as

ClathrinEndocytosisAlgorithmsAnimalsHumansMicroscopy, FluorescenceClathrinClathrin-mediated endocytosisColocalizationDrug-carrier internalizationDrug-carrier propertiesEndocytosisParticle trackingSmall unilamellar vesicle

Identifiers

PMID38971609

DOI10.1016/bs.mie.2024.02.010

PMCPMC11609598

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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.