This file ReadMe was updated on 2021-01-19 by Jay Werber ------------------- GENERAL INFORMATION ------------------- Title of Dataset: Supporting Data for Functionalized Polymersomes from a Polyisoprene-Activated Polyacrylamide Precursor Author Information: Principal Investigator Contact Information Name: Marc A. Hillmyer Institution: University of Minnesota Address: Chemistry, Minneapolis, Minnesota 55455 Email: hillmyer@umn.edu Associate or Co-investigator Contact information Name: Jay R. Werber Institution: University of Minnesota Address: Chemistry, Minneapolis, Minnesota 55455 Email: jwerber@umn.edu Associate or Co-investigator Contact Information Name: Colin Peterson Institution: University of Minnesota Address: Chemistry, Minneapolis, Minnesota 55455 Email: pet00842@umn.edu Associate or Co-investigator Contact Information Name: Nicholas J. Van Zee Institution: University of Minnesota Address: Chemistry, Minneapolis, Minnesota 55455 Email: vanze012@umn.edu Date of data collection (single date, range, approximate date): 2019-01-01 to 2020-09-30 Geographic location of data collection (where was data collected?): University of Minnesota Information about funding sources that supported the collection of the data: This research was supported by a grant from the U.S. Department of Energy, Officie of Science, Office of Basic Energy Sciences under Award Number DE-SC-0020210. -------------------------- SHARING/ACCESS INFORMATION -------------------------- 1. Licenses/restrictions placed on the data: CC0 1.0 Universal 2. Links to publications that cite or use the data: Werber, Jay R; Peterson, Colin; Van Zee, Nicholas J; Hillmyer, Marc A. Functionalized Polymersomes from a Polyisoprene-Activated Polyacrylamide Precursor. Langmuir 2021, 37 (1), 490. https://doi.org/10.1021/acs.langmuir.0c03157 3. Links to other publicly accessible locations of the data: N/A 4. Links/relationships to ancillary data sets: N/A 5. Was data derived from another source? No 6. Recommended citation for the data: Werber, Jay R; Peterson, Colin; Van Zee, Nicholas J; Hillmyer, Marc A. (2021). Supporting data for Functionalized Polymersomes from a Polyisoprene-Activated Polyacrylamide Precursor. Retrieved from the Data Repository for the University of Minnesota, https://doi.org/10.13020/k4rh-6j89. --------------------- DATA & FILE OVERVIEW --------------------- Purpose Statement: This document describes the raw datafiles used to generate the figures in the article "Functionalized Polymersomes from a Polyisoprene-Activated Polyacrylamide Precursor". General Notes: The files are organized by Figure and Scheme, with the type of analysis noted in the folder name and in the filenames. NMR files can be opened with MNOVA. SEC files are present as exported dRI data in Excel and as raw data files to be opened with Astra. IR files are present as raw data files to be opened in Opus. TEM files are tiff image files, and are the original files generated during electron microscopy. Confocal micoscopy files are ND2 files and exported JPG images of the same image. ND2 files are openable with Fiji. TGA files are present as TXT exports and raw data files to be opened with TA Universal Analysis. DSC files are present as XLS exports and .TRI files to be opened with TA Trios software. DLS files are present as .ARL files, which was the output of the REPES analysis. The .ARL files can be imported into Origin, and have two columns: 1) log(-gamma), from which radius can be calculated, and 2) the intensity-weighted prevalence. Are there multiple versions of the dataset? N -------------------------- METHODOLOGICAL INFORMATION -------------------------- Description of methods used for collection, generation, and processing of data: Nuclear magnetic resonance (NMR) spectroscopy was conducted on a Bruker Advance III HD 500 spectrometer. Chemical shifts are reported in δ units, expressed in ppm downfield of tetramethylsilane, using a residual CHCl3 peak as an internal standard (CDCl3, 1H: 7.26 ppm). Size exclusion chromatography (SEC) was conducted in THF at 25 °C, 1 mL/min, on an Agilent Infinity 1260 HPLC system equipped with three Waters Styragel HR columns in series, a Wyatt DAWN HELEOS-II 18-angle laser light scattering detector, and a Wyatt Optilab T-rEX differential refractive index detector. Polystyrene standards with conventional calibration were used to estimate molar mass. Thermogravimetric analysis (TGA) was conducted on a TA Instruments Q500. Differential scanning calorimetry (DSC) was conducted on a TA Instruments Discovery DSC using aluminum T-zero pans with hermetic lids. Infrared spectroscopy was conducted on a Bruker Alpha Platinum ATR spectrometer with a diamond ATR crystal. Cryo-electron microscopy (Cryo-EM) was used to image polymersomes, which were formed by film rehydration or direct dissolution in aqueous solutions. For film rehydration, a polymer was dissolved in THF or a 3:1 THF/MeOH mixture, with the solvent evaporated under a flow of N2 in a glass test tube, then under reduced pressure for ∼2 h. For film rehydration and direct dissolution of bulk polymer samples, deionized water or 1 mM tris(2-carboxyethyl)phosphine (TCEP)-HCl was added to the polymer to a concentration of 1−2 mg/mL, and the solution was bath-sonicated for 60 min and mixed by magnetic stirring for 1−3 days. The suspension was then extruded through polycarbonate track-etched membranes (Whatman) using a hand-held extruder (Avanti Polar Lipids). Vitrification of the polymersome solutions was carried out using an FEI Vitrobot Mark III automated vitrification device. Lacey carbon grids (200 mesh) were subjected to glow discharge to increase the surface energy of the grid. The grid was then transferred to an environmental chamber held at 100% humidity and 26 °C. The sample (6 μL) was then loaded onto the grid, blotted for 7 s, and then allowed to relax for 5 s before being plunged into a pool of liquid ethane. The grid was then stored in liquid nitrogen until imaging. Imaging was performed on an FEI Tecnai Spirit Bio-Twin TEM with an accelerating voltage of 120 kV. Samples were kept at a temperature of −177 ± 1 °C during imaging. Wall thicknesses were estimated using contrast intensity profiles in ImageJ, with thickness measured at 10 arbitrary locations per vesicle for 2−3 vesicles. The vesicle solutions assessed by Cryo-EM were analyzed for particle size distributions using dynamic light scattering (DLS), which was conducted with a Brookhaven BI-200SM instrument at 22 °C using a laser wavelength of 637 nm and a scattering angle of 90 °C. To obtain particle size distributions, the Laplace inversion routine REPES was used. Laser scanning confocal microscopy was conducted on a Nikon A1R fluorescence-lifetime imaging microscopy (FLIM) inverted confocal microscope with 488 and 561 nm laser excitation. Objectives used had 20× magnification immersed in air and 60× magnification immersed in oil. Dye-filled vesicles were prepared by film rehydration or direct dissolution in an aqueous buffer (5 mM Tris, 1 M NaCl, and 15 mM carboxyfluorescein, pH 7.5, or 10 mM Tris, 0.18 M NaCl, 10 mM carboxyfluorescein, and 1 mM TCEP, pH 7.5) and dialyzed for 1−3 days into an isosmotic buffer without carboxyfluorescein (5 mM Tris and 1.03 M NaCl, pH 7.5, or 10 mM Tris and 0.2 M NaCl, pH 7.5) using Spectrapor dialysis tubing with 12−14 kDa molecular weight cutoff (Spectrum Chemicals). Approximately 10 μL of the resulting solution was pipetted onto a glass microscope slide, covered with a coverslip, and sealed using clear nail polish. Images were analyzed in Fiji. People involved with sample collection, processing, analysis and/or submission: Jay Werber - Sample preparation, NMR, SEC, TGA, Confocal microscopy, DLS, IR Colin Peterson - DSC Nicholas Van Zee - Cryo-EM