Articles by Juan Pablo Giraldo in JoVE
Catalytic Scavenging of Plant Reactive Oxygen Species In Vivo by Anionic Cerium Oxide Nanoparticles Gregory Michael Newkirk*1,2, Honghong Wu*1, Israel Santana1, Juan Pablo Giraldo1,2 1Department of Botany and Plant Sciences, University of California, 2Department of Microbiology and Plant Pathology, University of California Here, we present a protocol for the synthesis and characterization of cerium oxide nanoparticles (nanoceria) for ROS (reactive oxygen species) scavenging in vivo, nanoceria imaging in plant tissues by confocal microscopy, and in vivo monitoring of nanoceria ROS scavenging by confocal microscopy.
Other articles by Juan Pablo Giraldo on PubMed
A Nanobionic Light-Emitting Plant Nano Letters. Dec, 2017 | Pubmed ID: 29148804 The engineering of living plants for visible light emission and sustainable illumination is compelling because plants possess independent energy generation and storage mechanisms and autonomous self-repair. Herein, we demonstrate a plant nanobionic approach that enables exceptional luminosity and lifetime utilizing four chemically interacting nanoparticles, including firefly luciferase conjugated silica (SNP-Luc), d-luciferin releasing poly(lactic-co-glycolic acid) (PLGA-LH), coenzyme A functionalized chitosan (CS-CoA) and semiconductor nanocrystal phosphors for longer wavelength modulation. An in vitro kinetic model incorporating the release rates of the nanoparticles is developed to maximize the chemiluminescent lifetimes to exceed 21.5 h. In watercress (Nasturtium officinale) and other species, the nanoparticles circumvent limitations such as luciferin toxicity above 400 μM and colocalization of enzymatic reactions near high adenosine triphosphate (ATP) production. Pressurized bath infusion of nanoparticles (PBIN) is introduced to deliver a mixture of nanoparticles to the entire living plant, well described using a nanofluidic mathematical model. We rationally design nanoparticle size and charge to control localization within distinct tissues compartments with 10 nm nanoparticles localizing within the leaf mesophyll and stomata guard cells, and those larger than 100 nm segregated in the leaf mesophyll. The results are mature watercress plants that emit greater than 1.44 × 10 photons/sec or 50% of 1 μW commercial luminescent diodes and modulate "off" and "on" states by chemical addition of dehydroluciferin and coenzyme A, respectively. We show that CdSe nanocrystals can shift the chemiluminescent emission to 760 nm enabling near-infrared (nIR) signaling. These results advance the viability of nanobionic plants as self-powered photonics, direct and indirect light sources.
Standoff Optical Glucose Sensing in Photosynthetic Organisms by a Quantum Dot Fluorescent Probe ACS Applied Materials & Interfaces. Aug, 2018 | Pubmed ID: 30058800 Glucose is a major product of photosynthesis and a key energy source for cellular respiration in organisms. Herein, we enable in vivo optical glucose sensing in wild-type plants using a quantum dot (QD) ratiometric approach. The optical probe is formed by a pair of QDs: thioglycolic acid-capped QDs which remain invariable to glucose (acting as an internal fluorescent reference control) and boronic acid (BA)-conjugated QDs (BA-QD) that quench their fluorescence in response to glucose. The fluorescence response of the QD probe is within the visible light window where photosynthetic tissues have a relatively low background. It is highly selective against other common sugars found in plants and can be used to quantify glucose levels above 500 μM in planta within the physiological range. We demonstrate that the QD fluorescent probe reports glucose from single chloroplast to algae cells ( Chara zeylanica) and plant leaf tissues ( Arabidopsis thaliana) in vivo via confocal microscopy and to a standoff Raspberry Pi camera setup. QD-based probes exhibit bright fluorescence, no photobleaching, tunable emission peak, and a size under plant cell wall porosity offering great potential for selective in vivo monitoring of glucose in photosynthetic organisms in situ.