Dimension 5

The θ–2θ diffraction data measurements are done on a Bruker D8 Discover diffractometer CuK_α1 (λ = 1.54 Å). Temperature-dependent electrical transport measurements are carried out in Physical Property Measurement system (Quantum Design) utilizing a four probe geometry. The surface morphology analysis of the various films are characterized on an atomic force microscope (AFM, Dimension V, Veeco).

Atomic force microscopy (AFM) measurements were carried out using a Veeco Dimension V instrument with aluminum-coated silicon AFM probes (resonant frequency 190 kHz). The system was operated in tapping mode with a VT-103-3K acoustic/vibration isolation system and a VT-102 vibration isolation table at room temperature in air. AFM was performed on Tissue Culture Polystyrene (TCPS) (Costar 3527, Corning, 24 well plate), glass H–ND monolayers. Scan sizes of 2 μm were taken and root mean square of the roughness (Rq) was calculated using Nanoscope Software 6.1.3. AFM Images were post processed with a median filter (3 × 3 kernel) using MATLAB 2012a software to remove noise and measurement artifacts.

Light microscopy was used to investigate the morphologies of obtained layers. For this purpose, we used IX71 (Olympus, Shinjuku, Japan) microscope operating in reflection mode and the Dimension V (Veeco, Plainview, New York, NY, USA) atomic force microscope (AFM) operating in tapping mode. The first technique was used to determine morphological changes at the micrometer-scale level. It provided a fast and rough estimation of samples morphologies. The insight into the morphology at the nanoscale level was assured by AFM microscopy. This technique allowed us to obtain images of high resolution, up to several nanometers; however, this technique is relatively slow and prone to distortions if scanning is fast and height amplitudes are significant. Both techniques are complementary to the research described in this article.
The thicknesses of layers were determined at the end of all measurements using a Dektak 3 (Veeco, Plainview, New York, NY, USA) profilometer. In this case, central regions of layers were removed using a scalpel to expose the surfaces of glass slides, and then the profilometer measured the depth of such a formed scratch.

To detect prostate microparticles in patient plasma samples, anti-PSMA mouse IgG_2b-PE (clone 3/E7) was added to 20μL of patient plasma. Negative isotype stained controls consisted of mouse isotype control IgG_2b-PE (Beckman Coutler Inc.) added to 20μL of patient plasma. Gates for each microparticle population were established by analyzing the isotype control first, modifying the gains for each PMT as necessary, and then analyzing the antibody labeled samples.
For microparticles generated in vitro, these were isolated by FACS as previously described in [12 (link)], in which PSMA+ve and zsGreen+ve co-expressing microparticle events were sorted onto cleaved mica coverslips (Ted Pella Inc. Redding, CA) to isolate prostate microparticles whilst eliminating non-target EVs and serum protein. After sorting 1000-3000 events onto the mica coverslips, samples were allowed to dry in 50 mL Falcon tubes for 5 minutes at 70°C before imaging with atomic force microscopy (AFM). No washing of the samples was performed. Images were acquired with tapping mode AFM (Dimension V, Veeco Inc.). Rectangle cantilevers with nominal spring constant of 40 N/m and tip radius of < 10 nm were used to acquire high resolution topographic images. All measurements were done at low driving force to avoid damaging samples.