Abstract：
:Carbon-based dots (CDs) and their functionalized (nano)composites have recently attracted attention due to their seemingly easy preparation and numerous potential applications, ranging from those in the biomedical field (i.e., imaging and drug delivery) to those in (opto)electronics (i.e., solar cells and LEDs). This protocol details step-by-step procedures for synthesis, purification, functionalization and characterization of nitrogen-doped carbon nanodots (NCNDs), which we have been preparing for the past few years. First, we describe the bottom-up synthesis of NCNDs, starting with the use of molecular precursors (arginine (Arg) and ethylenediamine (EDA)) and making use of microwave-assisted hydrothermal heating. We also provide guidelines for the purification of these materials, through either dialysis or low-pressure size-exclusion chromatography (SEC). Second, we outline post-functionalization procedures for the surface modification of NCNDs, such as alkylation and amidation reactions. Third, we provide instructions for the preparation of NCNDs with different properties, such as color emission, electrochemistry and chirality. Given the fast evolution of preparations and applications of CDs, issues that might arise from artifacts, errors and impurities should be avoided. In this context, the present protocol aims to provide details and guidelines for the synthesis of high-quality nanomaterials with high reproducibility, for various applications. Furthermore, specific needs might require the CDs to be prepared by different synthetic procedures and/or from different molecular precursors, but such CDs can still benefit from the purification and characterization procedures outlined in this protocol. The sample preparation takes various time frames, ranging from 4 to 18 d, depending on the adopted synthesis and purification steps.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Ðorđević L,Arcudi F,Prato Mdoi
10.1038/s41596-019-0207-xsubject
Has Abstractpub_date
2019-10-01 00:00:00pages
2931-2953issue
10eissn
1754-2189issn
1750-2799pii
10.1038/s41596-019-0207-xjournal_volume
14pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:High throughput microarray transcription analyses provide us with the expression profiles for large amounts of plant genes. However, their tissue and cellular resolution is limited. Thus, for detailed functional analysis, it is still necessary to examine the expression pattern of selected candidate genes at a cellular...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.333
更新日期：2006-01-01 00:00:00
abstract：:This protocol describes an optimized method for direct in vitro monitoring of homo- and heterotypic axon-axon interactions involved in the developmental assembly of neural circuits. The assay exploits a classical example of heterotypic axonal interactions by modeling the sequential extension of spinal motor and somato...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.442
更新日期：2012-01-26 00:00:00
abstract：:Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent c...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.156
更新日期：2013-01-01 00:00:00
abstract：:The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabricati...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.027
更新日期：2012-04-05 00:00:00
abstract：:This protocol represents a novel enzyme-luminescence method to detect dopamine sensitively and rapidly with high temporal resolution. In principle, dopamine is first oxidized with tyramine oxidase to produce H(2)O(2), and then the produced H(2)O(2) reacts with luminol to generate chemiluminescence in the presence of h...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.158
更新日期：2008-01-01 00:00:00
abstract：:Venous access catheters used in clinics are prone to biofilm contamination, contributing to chronic and nosocomial infections. Although several animal models for studying device-associated biofilms were previously described, only a few detailed protocols are currently available. Here we provide a protocol using totall...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.033
更新日期：2016-03-01 00:00:00
abstract：:Studies on colonic cells in the lamina propria (LP) of mice are important for understanding the cellular and immune responses in the gut, especially in inflammatory bowel diseases (such as morbus crohn and colitis ulcerosa). This protocol details a method to isolate LP cells and characterize freshly isolated cells by ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.315
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled micros...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.307
更新日期：2011-05-01 00:00:00
abstract：:This protocol describes a method for direct labeling and detection of small RNAs present in total RNA by splinted ligation. The assay uses a small RNA-specific bridge oligonucleotide to form base pairs with the small RNA and a 5'-end-radiolabeled ligation oligonucleotide. The captured small RNA is directly labeled by ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.530
更新日期：2008-01-01 00:00:00
abstract：:Cell migration is a key feature of virtually every biological process, and it can be studied in a variety of ways. Here we outline a protocol for the in vitro study of cell migration using a ring barrier-based assay. A 'barrier' is inserted in the culture chamber, which prevents cells from entering a defined area. Cel...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.056
更新日期：2015-06-01 00:00:00
abstract：:Cap-analysis gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. CAGE allows mapping of all the initiation sites of both capped coding and noncoding RNAs. In addition, transcriptional start sites within promoters are characterized at single-nucleotide resolution. The latter allows t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.005
更新日期：2012-02-23 00:00:00
abstract：:At present, several assays that use radioisotope labeling to quantify the degradation of long-lived proteins have been developed to measure autophagic flux. Here, we describe a nonradioactive pulse-chase protocol using L-azidohomoalanine (AHA) labeling to quantify long-lived protein degradation during autophagy. AHA i...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.160
更新日期：2017-12-01 00:00:00
abstract：:Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.17
更新日期：2006-01-01 00:00:00
abstract：:Systematic complex genetic interaction studies have provided insight into high-order functional redundancies and genetic network wiring of the cell. Here, we describe a method for screening and quantifying trigenic interactions from ordered arrays of yeast strains grown on agar plates as individual colonies. The proto...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-020-00456-3
更新日期：2021-01-18 00:00:00
abstract：:The direct detection of drug-protein interactions in living cells is a major challenge in drug discovery research. Recently, we introduced an approach termed thermal proteome profiling (TPP), which enables the monitoring of changes in protein thermal stability across the proteome using quantitative mass spectrometry. ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.101
更新日期：2015-10-01 00:00:00
abstract：:The research community is in a race to understand the molecular mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, to repurpose currently available antiviral drugs and to develop new therapies and vaccines against coronavirus disease 2019 (COVID-19). One major challenge in achieving ...
journal_title：Nature protocols
pub_type： 杂志文章,评审
doi：10.1038/s41596-020-00403-2
更新日期：2020-12-01 00:00:00
abstract：:Hepatic stellate cells (HSCs) have been identified as the main fibrogenic cell type in the liver. Hence, efforts to understand hepatic fibrogenesis and to develop treatment strategies have focused on this cell type. HSC isolation, originally developed in rats, has subsequently been adapted to mice, thus allowing the s...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.017
更新日期：2015-02-01 00:00:00
abstract：:Reversible thiol oxidation of cysteine residues occurs in many intracellular catalytic and signaling processes. Here we describe an optimized protocol, which can be completed in ∼5 d, to unambiguously identify specific cysteine residues that are transiently and reversibly oxidized by comparing two complex biological s...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.065
更新日期：2014-05-01 00:00:00
abstract：:This transformation procedure generates, with high efficiency (70-90%), hairy roots in cultivars, landraces and accessions of Phaseolus vulgaris (common bean) and other Phaseolus spp. Hairy roots rapidly develop after wounding young plantlets with Agrobacterium rhizogenes, at the cotyledon node, and keeping the plants...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.259
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes a simple means of measuring the starch content of plant tissues by solubilizing the starch, converting it quantitatively to glucose and assaying the glucose. Plant tissue must initially be frozen rapidly to stop metabolism, then extracted to remove free glucose. Starch is solubilized by heating...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.232
更新日期：2006-01-01 00:00:00
abstract：:Surface- and matrix-bound signals modulate stem cell fate in vivo and in vitro. This protocol enables the immobilization of a wide range of biomolecules that contain primary amino groups to different types of solid carriers, including glass substrates and standard polystyrene well plates. We describe how thin polymer ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.70
更新日期：2010-06-01 00:00:00
abstract：:This protocol describes a directed evolution method for in vitro mutagenesis and recombination of polynucleotide sequences. The staggered extension process (StEP) is essentially a modified PCR that uses highly abbreviated annealing and extension steps to generate staggered DNA fragments and promote crossover events al...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.309
更新日期：2006-01-01 00:00:00
abstract：:This protocol describes a methodology for imaging the sequestration of infected erythrocytes of the rodent malaria parasite Plasmodium berghei in the bodies of live mice or in dissected organs, using a transgenic parasite that expresses luciferase. Real-time imaging of infected erythrocytes is performed by measuring b...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.69
更新日期：2006-01-01 00:00:00
abstract：:Neurons in cortical sensory regions receive modality-specific information through synapses that are located on their dendrites. Recently, the use of two-photon microscopy combined with whole-cell recordings has helped to identify visually evoked dendritic calcium signals in mouse visual cortical neurons in vivo. The c...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.169
更新日期：2011-01-01 00:00:00
abstract：:The synthesis of an azobenzene amino acid (aa) for use as a photo-inducible conformational switch in polypeptides is described. The compound can be easily incorporated into an aa sequence by solid-phase peptide synthesis using standard 9-fluorenylmethoxycarbonyl methods. A reversible conformational change of the pepti...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.488
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes a simple silver staining method used to visualize DNA fragments and other organic molecules with unsurpassed detail following traditional polyacrylamide gel electrophoresis (PAGE). Sensitivity rivals radioisotopic methods and DNA in the picogram range can be reliably detected. The described pro...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.330
更新日期：2007-01-01 00:00:00
abstract：:This protocol describes the efficient, generally applicable Ullmann coupling reaction of bromaminic acid with alkyl- or aryl-amines in phosphate buffer under microwave irradiation using elemental copper as a catalyst. The reaction leads to a number of biologically active compounds. As a prototypical example, the synth...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.63
更新日期：2010-05-01 00:00:00
abstract：:The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its invo...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.082
更新日期：2016-09-01 00:00:00
abstract：:This protocol describes a simple but robust microfluidic assay combining three-dimensional (3D) and two-dimensional (2D) cell culture. The microfluidic platform comprises hydrogel-incorporating chambers between surface-accessible microchannels. By using this platform, well-defined biochemical and biophysical stimuli c...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.051
更新日期：2012-06-07 00:00:00
abstract：:Transitivity Clustering is a method for the partitioning of biological data into groups of similar objects, such as genes, for instance. It provides integrated access to various functions addressing each step of a typical cluster analysis. To facilitate this, Transitivity Clustering is accessible online and offers thr...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2010.197
更新日期：2011-03-01 00:00:00
