Analytical methods for copper, zinc and iron quantification in mammalian cells

Cerchiaro, Giselle; Manieri, Tânia Maria; Bertuchi, Fernanda R.

doi:10.1039/c3mt00136a

Cited by 61 publications

(42 citation statements)

References 106 publications

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“…A modest response is observed with free copper salts, as is similarly observed for the related fluorescence probe FIP-1 (15). However, as a typical eukaryotic cell exhibits a ∼10-fold higher level of iron over copper coupled with the high buffering capacity of copper with glutathione and metallochaperones (picomolar to femtomolar K d values) (55)(56)(57)(58)(59)(60), the modest response to free copper salts suggests that ICL-1 should have sufficient selectivity to detect alterations in biological ferrous iron levels. ICL-1 is also selective for labile Fe 2+ over other biologically relevant forms of iron that are tightly bound to proteins and cofactors, such as transferrin, ferritin, hemin, and hemoglobin, as well as Fe 3+ , along with reductants glutathione, N-acetyl cysteine, β-mercaptoethanol, and ascorbic acid (Fig.…”

Section: Resultsmentioning

confidence: 90%

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

Aron

Heffern

Lonergan

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

113

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Iron is an essential metal for all organisms, yet disruption of its homeostasis, particularly in labile forms that can contribute to oxidative stress, is connected to diseases ranging from infection to cancer to neurodegeneration. Iron deficiency is also among the most common nutritional deficiencies worldwide. To advance studies of iron in healthy and disease states, we now report the synthesis and characterization of iron-caged luciferin-1 (ICL-1), a bioluminescent probe that enables longitudinal monitoring of labile iron pools (LIPs) in living animals. ICL-1 utilizes a bioinspired endoperoxide trigger to release D-aminoluciferin for selective reactivity-based detection of Fe 2+ with metal and oxidation state specificity. The probe can detect physiological changes in labile Fe 2+ levels in live cells and mice experiencing iron deficiency or overload. Application of ICL-1 in a model of systemic bacterial infection reveals increased iron accumulation in infected tissues that accompany transcriptional changes consistent with elevations in both iron acquisition and retention. The ability to assess iron status in living animals provides a powerful technology for studying the contributions of iron metabolism to physiology and pathology.labile iron | molecular imaging | luciferin | metal homeostasis | infectious disease I ron is an essential mineral for nearly every form of life, owing in large part to its ability to cycle between different oxidation states for processes such as nucleotide synthesis, oxygen transport, and respiration (1, 2). At the same time, the potent redox activity of iron is potentially toxic, particularly in unregulated labile forms that can trigger aberrant production of reactive oxygen species via Fenton chemistry (3). Indeed, iron deficiency remains one of the most common nutritional deficiencies in the world (4), and aberrant iron levels have been linked to various ailments, including cancer (5-7), cardiovascular (8), and neurodegenerative (9) disorders, as well as aging (10). The situation is especially complex in infectious diseases, where the requirement for iron by both host organism and invading pathogen leads to an intricate chemical tug-of-war for this metal nutrient during various stages of the immune response (11,12).The foregoing examples provide motivation for developing technologies to monitor biological iron status, with particular interest in methods to achieve in vivo iron imaging in live animal models that go beyond current state-of-the-art assays that are limited primarily to cell culture specimens. In this regard, detection of iron with both metal and oxidation state specificity is of central importance, because while iron is stored primarily in the ferric oxidation state, a ferrous iron pool loosely bound to cellular ligands, defined as the labile iron pool (LIP), exists at the center of highly regulated networks that control iron acquisition, trafficking, and excretion. Indeed, as a weak binder on the Irving-Williams stability series (13), Fe 2+ provides a challenge for d...

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Section: Resultsmentioning

confidence: 90%

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

Aron

Heffern

Lonergan

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

113

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“…[1] The utility of metal ions cannot be ascertained unless we are able to quantify the metal content of cells and tissues. Previously, the quantification of the metal ions was done via Atom Absorption Spectroscopy [2][3][4] and volumetric detection. [5] With technological advancement, these studies subsequently gave way to Inductively Coupled Plasma Mass Spectrometry (ICP-MS), Inductively Coupled Plasma Atomic Emission Spectrometry (ICP-AES) and preconcentration techniques involving solid-phase extraction of heavy metal ions as analytical methods owing to their superior sensitivity.…”

Section: Introductionmentioning

confidence: 99%

Water-soluble polymeric chemosensor for detection of Cu²⁺ions with high selectivity and sensitivity

Maiti

Banerjee

Maiti

et al. 2016

Designed Monomers and Polymers

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Development of water-soluble chemosensors that are selective and sensitive to Cu 2+ ions is of tremendous importance owing to their potential applications in biological systems. In the present work, we report the synthesis of a new water-soluble polymer containing pendant rhodamine units that are capable of highly selective and sensitive detection of Cu 2+ ions in aqueous medium. Poly(2-pyrrolidinemethyl acrylate) was prepared using RAFT polymerization technique. The pyrrolidine nitrogen group in the polymer was subjected to Aza-Michael type addition with ethyl acrylate that was followed by covalent linking of rhodamine units to the polymer. This polymer was completely water-soluble and found to be capable of sensing Cu 2+ ions in aqueous medium. Cu 2+ -induced opening of the spirolactam ring of the rhodamine units resulted in rapid and easily noticeable colour change, thus enabling a highly selective detection of Cu 2+ in μmol range. The ability of these polymeric systems to detect Cu 2+ ions in complete aqueous media has more importance than use of organic solvents to solubilize the polymer as reported previously, and thus opened a new window for application of these systems in the detection of copper ions in biological systems.

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“…The attachment of AgNP-aggregates to single bacterial cells was identified by TEM and SEM-EDX (Guo et al, 2017). Aside from TEM and SEM-EDX, other techniques have been used to visualize and identify metals in single human cells, e.g., AFM, SXFM (Cerchiaro et al, 2013). However, these microscopy techniques have limitations including random sample detection, and lack of quantitative information regarding cellular interactions with nanoparticles.…”

Section: Introductionmentioning

confidence: 99%

Mass Cytometry for Detection of Silver at the Bacterial Single Cell Level

et al. 2017

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Background: Mass cytometry (Cytometry by Time of Flight, CyTOF) allows single-cell characterization on the basis of specific metal-based cell markers. In addition, other metals in the mass range such as silver can be detected per cell. Bacteria are known to be sensible to silver and a protocol was developed to measure both the number of affected cells per population and the quantities of silver per cell.Methods: For mass cytometry ruthenium red was used as a marker for all cells of a population while parallel application of cisplatin discriminated live from dead cells. Silver quantities per cell and frequencies of silver containing cells in a population were measured by mass cytometry. In addition, live/dead subpopulations were analyzed by flow cytometry and distinguished by cell sorting based on ruthenium red and propidium iodide double staining. Verification of the cells’ silver load was performed on the bulk level by using ICP-MS in combination with cell sorting. The protocol was developed by conveying both, fast and non-growing Pseudomonas putida cells as test organisms.Results: A workflow for labeling bacteria in order to be analyzed by mass cytometry was developed. Three different parameters were tested: ruthenium red provided counts for all bacterial cells in a population while consecutively applied cisplatin marked the frequency of dead cells. Apparent population heterogeneity was detected by different frequencies of silver containing cells. Silver quantities per cell were also well measurable. Generally, AgNP-10 treatment caused higher frequencies of dead cells, higher frequencies of silver containing cells and higher per-cell silver quantities. Due to an assumed chemical equilibrium of free and bound silver ions live and dead cells were associated with silver in equal quantities and this preferably during exponential growth. With ICP-MS up to 1.5 fg silver per bacterial cell were detected.Conclusion: An effective mass cytometry protocol was developed for the detection and quantification of silver in single bacterial cells of different physiological states. The silver quantities were generally heterogeneously distributed among cells in a population, the degree of which was dependent on micro-environmental conditions and on silver applied either in ion or nanoparticle-aggregated form.

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Analytical methods for copper, zinc and iron quantification in mammalian cells

Cited by 61 publications

References 106 publications

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

Water-soluble polymeric chemosensor for detection of Cu²⁺ions with high selectivity and sensitivity

Mass Cytometry for Detection of Silver at the Bacterial Single Cell Level

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Analytical methods for copper, zinc and iron quantification in mammalian cells

Cited by 61 publications

References 106 publications

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

In vivo bioluminescence imaging of labile iron accumulation in a murine model of Acinetobacter baumannii infection

Water-soluble polymeric chemosensor for detection of Cu2+ions with high selectivity and sensitivity

Mass Cytometry for Detection of Silver at the Bacterial Single Cell Level

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Product

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Water-soluble polymeric chemosensor for detection of Cu²⁺ions with high selectivity and sensitivity