Digital micrograph imaging software

NP size was measured by dynamic light scattering (DLS) using a Zetasizer Nano apparatus (Malvern, UK). The NP was imaged under transmission electron microscope (TEM) (JEM-2200FM, Jeol Ltd., Akishima, Tokyo, Japan) operating at 200 kV using a Gatan multiscan CCD camera with Digital Micrograph imaging software (Gatan, Pleasanton, CA, USA). Uranyl acetate staining was employed for TEM imaging. UV spectrophotometry was used to quantify the concentration of ATS9R and ASO within the NP complex. Individual standard curves of ATS9R and ASO in solution were created to quantify samples with an absorbance at 260 nm indicative of the ASO and that at 450 nm indicative of the ATS9R peptide. Using these values, the absorbance values at 260 nm and 450 nm of the NP complex could be used to calculate the relative ratio of ASO and ATS9R in the NP based on the respective standard curves based on the Beer−Lambert law. When a 1:1 ASO and ATS9R peptide were mixed together, the molar ratio of ASO and ATS9R in the resulting NP was determined to be 1:1 from the absorbance values.

Visualization of the bacteriophage phenotype was achieved using a JEOL JEM-2100 transmission electron microscope (TEM) operated at 200 kV as previously described (Kabwe et al., 2020 (link)). Using a 400-mesh formvar and carbon copper grids (ProScieTech, Australia), EFA1 bacteriophage particles were adsorbed for 1 min before rinsing the grids with milli-Q water and negatively staining for 20 s with 2% (w/v) uranyl acetate (Sigma-Aldrich^®, Australia). Excess uranyl acetate was removed, and the copper grids allowed to air dry at room temperature for 30 min. The Gatan Orius SC200D 1 wide-angle camera coupled to the Gatan Microscopy Suite and Digital Micrograph Imaging software (Version 2.3.2.888.0) was used to take the TEM images before being exported to Image J (Version 1.8.0_112) for further analysis.

Liquid
cells were prepared
as previously described in the literature.^{71 (link)} Briefly, solutions of ethanol and trans-anethole
were drop cast onto nonglow discharged SiN_x chips in amounts less than 0.8 μL. Top chips were deposited
such that the windows were aligned orthogonally, and the holder was
sealed with the top clamp assembly. Lines were left unfilled with
solvent so as to avoid premature dilution of the sample with diluent.
The holder was then pumped down using the external pumping station,
and the cell windows were visually inspected using the attached optical
microscope. Once the cell has reached 8.6 × 10^–6 mbar, the fluidic ports were unsealed and the flow line was attached,
so as to ensure cell integrity during dilution. This flow line was
attached to a syringe and syringe pump, which were used to flow in
water to the cell at rates from 1 to 5 μL/min. A JEM-ARM300F
transmission electron microscope operating at a voltage of 300 keV
and current of 15 μA (FEG source) was used for liquid cell experiments.
Images were acquired via Gatan 2k × 2k OneView IS CMOS camera via Gatan Digital Micrograph imaging
software with exposures of 1 s.