Ultrascan 2k 2k ccd

An aliquot of Pro-drug-NP and Lipid-NP was drop-casted onto a carbon coated copper grid (200 mesh size). Lipid-NP samples were stained with 10 μl of uranyl acetate (1%). TEM images were taken on JEOL 2100 Cryo TEM machine and imaged by Gatan UltraScan 2k × 2k CCD. The concentrations were used as Pro-drug-NP containing 400 μM of Pro-Drug molecules and equivalents of LysoPC29 .

DCA, MCA, DBA were obtained from Sigma Aldrich, Inc. (St. Louis, MO) while LysoPC was purchased from Avanti Polar Lipids (Birmingham, Al). The hydrodynamic diameter was measured on a Malvern Zetasizer machine equipped with 633 nm laser. Zeta potential measurement was performed on a Malvern Zetasizer instrument, from MRL facility, UIUC. The TEM images were acquired on a JEOL 2100 Cryo TEM machine and imaged by Gatan UltraScan 2k × 2k CCD. The XRD data was collected on instrument Siemens-Bruker D5000 diffractometer and analyzed using software Jade X-ray analysis. MTT reduction assay was ended with performing absorption assay on plate reader (Synergy HT, Bio-Tek). Bright field imaging was performed on microscope DMI3000 B, Leica Microsystems, Buffalo Grove, IL.

For TEM, Au-PEG clusters were diluted 100-fold with carbon-filtered deionized water (0.2 μm cellulosic membrane, pH = 7) and 2.5 μl of the diluted Au-PEG clusters were placed on a 300-mesh carbon film supported by a copper grid and allowed to stabilize for 2 min. A filter paper was then used to remove liquid for thin film formation and then allowed to air dry while covered. Images were obtained using a Jeol 2010 cryo-electron microscope operating at 200 kV, and using different degrees of defocus to obtain an adequate bright contrast. Images were recorded on a Gatan UltraScan 2k × 2k CCD. These CCD images were processed and analyzed with ImageJ (http://rsbweb.nih.gov/ij/) version 1.48.