By Daniel F. Moyano, Vincent M. Rotello (auth.), Volkmar Weissig, Tamer Elbayoumi, Mark Olsen (eds.)
In Cellular and Subcellular Nanotechnology: equipment and Protocols expert researchers within the box element the newest advances which were made in using the large capability of nanotechnology for probing, imaging and manipulating lifestyles on a mobile and subcellular point. Written within the hugely winning Methods in Molecular Biology™ sequence structure, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, effortlessly reproducible laboratory protocols, and key pointers on troubleshooting and fending off recognized pitfalls.
Authoritative and useful, Cellular and Subcellular Nanotechnology: equipment and Protocols seeks to assist scientists within the extra examine of utilising nanotechnology to all parts of biomedical sciences.
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Extra info for Cellular and Subcellular Nanotechnology: Methods and Protocols
3). 38 Valeriy Lukyanenko and Vadim Salnikov Fig. 3 Stabilizing the effect of polymer polyvinylpyrrolidone (PVP) on gold nanoparticles. 2), and (c) in particles pretreated with 1% PVP and in aspartic acid solution. The suspensions were airdried on formvar-coated grids. No staining. (d) The absorption spectra for colloidal gold (peak at 520 nm) and mixtures of gold and 150 mM potassium aspartate and/or 5% PVP. OD optical density (depends on gold particles concentration); dashed lines show the absorbance maximum for colloidal gold (523 nm).
3. Dounce homogenizer (7 mL). 4. 15 mL centrifuge tubes. 5. Refrigerated centrifuge. 4 Isolation of NanoparticleContaining Endosomes 1. 3). 2. 1). nanoparticle-treated with attached 3. Magnetic separator rack. 4. Sterile PBS. 5. Refrigerated tabletop centrifuge. 5 Quantification of Drug Content by HPLC 43 1. HPLC-grade methanol. 2. Sonicator disruptor with probe tip. 3. Refrigerated tabletop centrifuge. 4. 5 mL microcentrifuge tubes. 5. HPLC autoinjector vials with low-volume inserts. 1 Conjugate Antibodies to Magnetic Beads 1.
Dry them. Now the slices are ready for electron microscopy. 11. Store images in tiff format (see Notes 3 and 4). Representative micrographs (Fig. 2) show the typical distribution of nanoparticles within ventricular cardiomyocytes. 4 Notes 1. To prevent aggregation of gold nanoparticles and their binding to proteins, after conjugation, gold sols have to be coated with polyvinylpyrrolidone (PVP) or polyethylene glycol PEG (Fig. 3). 38 Valeriy Lukyanenko and Vadim Salnikov Fig. 3 Stabilizing the effect of polymer polyvinylpyrrolidone (PVP) on gold nanoparticles.