Data from: Mechanical performance of hybrid polymer-lipid vesicles with leaflet asymmetry engineered using microfluidics
Data files
Mar 07, 2026 version files 57.52 MB
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pnas_dataverse_01232026.zip
57.51 MB
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README.md
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Abstract
Lipid vesicles consist of aqueous cores surrounded by a bilayer of phospholipids. Hybrid polymer–lipid vesicles incorporate both polymers and lipids, offering promising properties for developing pharmaceuticals, biosensors, and artificial cells. The hybrid vesicles can be symmetric, with two leaflets of identical compositions, or asymmetric, with leaflets of dissimilar compositions, which can lead to dramatically altered properties. However, existing methods for producing symmetric and asymmetric hybrid vesicles often result in heterogeneous compositions and sizes, making it challenging to quantify the effect of asymmetry and limiting applications. Here, we use a microfluidic approach to produce hybrid vesicles with either symmetric or asymmetric leaflets and precisely engineered compositions. We find that the vesicles with asymmetric leaflets are significantly stiffer and tougher than those with symmetric leaflets; moreover, the lateral diffusivity of lipids is greatly decreased. The structure for improved toughness consists of a stretchable lipid inner leaflet and a fully continuous polymer outer leaflet. This approach to precisely engineer asymmetric structures can be applied to hybrid vesicles composed of block copolymers and phospholipids soluble in chloroform and hexane, further expanding their applications.
Dataset DOI: 10.5061/dryad.cjsxksnmm
README: Mechanical performance of hybrid polymer–lipid vesicles with leaflet asymmetry engineered using microfluidics
These data support the findings of the article:
Huang Y., Manafirad A., Matoori S., Arriaga L.R., Sun S., Chen A., Dinsmore A.D., Mooney D.J., Weitz D.A.
“Mechanical performance of hybrid polymer–lipid vesicles with leaflet asymmetry engineered using microfluidics”
Proceedings of the National Academy of Sciences (PNAS).
Article link:
https://www.pnas.org/doi/10.1073/pnas.2516407123\
Description of the data
This dataset (pnas_dataverse_01232026.zip) contains processed measurements used to generate the figures and statistical analyses reported in the study. The data include vesicle size measurements, micropipette aspiration mechanical measurements, and statistical summaries of vesicle properties.
DATASET ORGANIZATION
Figure1_Vesicle_Image_Analysis
This folder contains representative microscopy images and MATLAB scripts used for vesicle contour detection and image-based size analysis, as well as example images illustrating vesicle dewetting and phase separation phenomena described in the manuscript.
Contour Analysis
The folder "contour analysis" contains example microscopy images and MATLAB scripts for detecting vesicle contours and quantifying vesicle size distributions.
Example image files:
example_12_01212026.png
example_15.png
example_16.png
These images show representative vesicles used during the development and validation of the contour detection workflow.
Detection output images:
example_15_detection.png
example_16_detection.png
These images show the vesicle contour detection results produced by the MATLAB image analysis pipeline.
CV output images:
Example_12_CV.png
example_15_CV.png
example_16_CV.png
These figures show intermediate image-processing outputs used for contour extraction and verification of vesicle detection.
MATLAB scripts:
step1_sample01222026_example_12_main.m – Main MATLAB script used to perform contour-based vesicle detection and measurement from microscopy images.
threshold_circles.m – MATLAB function used to detect circular vesicle boundaries based on intensity thresholding.
threshold_circles_li.m – Alternative threshold-based circle detection function used for vesicle segmentation.
Dewetting
The folder "dewetting" contains representative microscopy images showing vesicle dewetting behaviour observed during experiments.
dewetting1.png
dewetting2.png
These images illustrate morphological changes in hybrid vesicles consistent with dewetting phenomena described in the manuscript.
Phase Separation
The folder "phase separation" contains representative microscopy images showing phase-separated membrane domains in hybrid polymer–lipid vesicles.
phase separation 1
phase separation 2
phase separation 3
phase separation 4
phase separation 5
These images illustrate examples of membrane phase separation observed in vesicles with different polymer–lipid compositions.
Figure2_Asymmetric_Vesicle_Production
Representative videos documenting the formation of asymmetric vesicles using the microfluidic device described in the manuscript.
Files included:
• vesicle_generation_video1.mp4 – Microfluidic experiment showing vesicle formation.
• vesicle_generation_video2.mp4 – Example showing stable droplet formation during vesicle production.
These videos illustrate the hydrodynamic conditions used to control vesicle diameter.
Variables
No numerical variables are associated with these files; they serve as qualitative visual documentation of vesicle production.
Figure3_Micropipette_Aspiration
This folder contains summary statistics derived from micropipette aspiration experiments used to determine the mechanical properties of vesicles. The spreadsheets include calculated quantities such as membrane tension, maximum strain, stretching modulus (slope of the tension–strain curve), and toughness. These values correspond to the vesicle types analysed in the manuscript.
Excel data files
- max_strain.xlsx – Summary statistics of the maximum areal strain reached by vesicles during micropipette aspiration experiments.
- max_tau.xlsx – Summary statistics of the maximum membrane tension measured for each vesicle type.
- min_tau.xlsx – Summary statistics of the minimum membrane tension values.
- slope_m.xlsx – Mean values of the slope of the tension–strain relationship (stretching modulus).
- slope_err.xlsx – Standard errors associated with the slope measurements.
- tough_m.xlsx – Mean values of membrane toughness calculated from the tension–strain curves.
- tough_err.xlsx – Standard errors associated with toughness measurements.
- maxs_m.xlsx, maxs_err.xlsx – Mean values and associated errors for maximum strain measurements.
- maxtau_m.xlsx, maxtau_err.xlsx – Mean values and associated errors for maximum membrane tension.
- mintau_m.xlsx, mintau_err.xlsx – Mean values and associated errors for minimum membrane tension.
- slope_mat.xlsx – Matrix containing slope values for each vesicle group used to calculate statistical summaries.
- total.xlsx – Combined summary statistics used to generate the figures in the manuscript.
MATLAB scripts
- paper_statistics_step1.m – MATLAB script used to process micropipette aspiration measurements and compute statistical summaries.
- paper_statistics_step2.m – MATLAB script used to calculate group statistics and prepare data for plotting.
- protocol_bar_error_2Groups.m – MATLAB script used to generate bar plots with error bars.
- protocol_errorPlot.m – MATLAB function used to generate statistical error plots.
- protocol_figure_format.m, protocol_figure_format_3.m – MATLAB scripts used to format figures for the manuscript.
- paper_protocol_figure_format_simple1_step3.m – MATLAB script used to generate final formatted figures.
Figure files
- paper_maxtau_tough.eps – Figure illustrating maximum tension and toughness results.
- paper_stretching_modulus_lysis_strain.eps – Figure illustrating stretching modulus and lysis strain results.
- Figure4_FRAP_Analysis
This folder contains representative fluorescence recovery after photobleaching (FRAP) experiments used to quantify membrane mobility in hybrid polymer–lipid vesicles. The folder includes representative videos of bleached vesicle regions, processed FRAP datasets, and MATLAB scripts used for analysis and figure generation.
FRAP Representative Videos
The folder "FRAP representative videos of different bleached spots" contains example recordings of vesicles after photobleaching.
large.avi – FRAP experiment with a large bleached region.
medium.avi – FRAP experiment with a medium-sized bleached region.
small.avi – FRAP experiment with a small bleached region.
These videos illustrate fluorescence recovery dynamics for vesicles with different bleaching spot sizes.
FRAP Statistics Summary
This folder contains CSV files with fluorescence intensity measurements extracted from FRAP experiments for different vesicle compositions.
Example datasets include:
08012024_asy2_x_4halfLw2_y_w2.csv
asy_dopc_in_polymer_out_x_4halfLw2_y_w2_07262024_3.csv
asy2_x_4halfLw2_y_w2.csv
paper_08012024_2_asy2_x_4halfLw2_y_w2.csv
paper_asy_x_4halfLw2_y_w2_07262024_4.csv
paper_DOPC_x_4halfLw2_y_w2_07262024_2.csv
paper_dopc20_polymer80_x_4halfLw2_y_w2_07262024_3.csv
paper_dopc80_polymer20_x_4halfLw2_y_w2_07262024_2.csv
Each CSV file contains processed FRAP measurements for a specific vesicle composition.
Variables in FRAP CSV files
- Time_s – Time after photobleaching (seconds).
- Fluorescence_Intensity – Measured fluorescence intensity within the bleached region.
- Normalized_Intensity – Fluorescence intensity normalised to the pre-bleach signal.
MATLAB Analysis Files
figure_format_3.m – MATLAB script used to format FRAP plots for publication figures.
paper_frap_slope_D.m – MATLAB script used to calculate fluorescence recovery slopes and diffusion-related metrics from FRAP data.
protocol_errorPlot.m – MATLAB function used to generate plots with statistical error bars.
Additional Files
matlab_workspace.mat – MATLAB workspace file containing intermediate analysis variables used during FRAP data processing.
Software Requirements
The analysis scripts were written in MATLAB and tested using MATLAB R2022b.
The scripts use standard MATLAB functions and do not require additional toolboxes.
Reproducibility
All MATLAB scripts included in this repository reproduce the quantitative analyses reported in the manuscript. Scripts are organized according to the corresponding figure and operate directly on the datasets included in the repository.
Contact Information
Questions regarding the dataset or analysis scripts can be directed to the corresponding author listed in the manuscript.