An approach to estimate spatial distribution of analyte within cells using spectrally-resolved fluorescence microscopy.

While fluorescence microscopy has become an essential tool amongst chemists and biologists for the detection of various analyte within cellular environments, non-uniform spatial distribution of sensors within cells often restricts extraction of reliable information on relative abundance of analytes in different subcellular regions. As an alternative to existing sensing methodologies such as ratiometric or FRET imaging, where relative proportion of analyte with respect to the sensor can be obtained within cells, we propose a methodology using spectrally-resolved fluorescence microscopy, via which both the relative abundance of sensor as well as their relative proportion with respect to the analyte can be simultaneously extracted for local subcellular regions. This method is exemplified using a BODIPY sensor, capable of detecting mercury ions within cellular environments, characterized by spectral blue-shift and concurrent enhancement of emission intensity. Spectral emission envelopes collected from sub-microscopic regions allowed us to compare the shift in transition energies as well as integrated emission intensities within various intracellular regions. Construction of a 2D scatter plot using spectral shifts and emission intensities, which depend on the relative amount of analyte with respect to sensor and the approximate local amounts of the probe, respectively, enabled qualitative extraction of relative abundance of analyte in various local regions within a single cell as well as amongst different cells. Although the comparisons remain semi-quantitative, this approach involving analysis of multiple spectral parameters opens up an alternative way to extract spatial distribution of analyte in heterogeneous systems. The proposed method would be especially relevant for fluorescent probes that undergo relatively nominal shift in transition energies compared to their emission bandwidths, which often restricts their usage for quantitative ratiometric imaging in cellular media due to strong cross-talk between energetically separated detection channels.

Infinite Polyiodide Chains in the Pyrroloperylene-Iodine Complex: Insights into the Starch-Iodine and Perylene-Iodine Complexes.

We report the preparation and X-ray crystallographic characterization of the first crystalline homoatomic polymer chain, which is part of a semiconducting pyrroloperylene-iodine complex. The crystal structure contains infinite polyiodide I∞ (δ-) . Interestingly, the structure of iodine within the insoluble, blue starch-iodine complex has long remained elusive, but has been speculated as having infinite chains of iodine. Close similarities in the low-wavenumber Raman spectra of the title compound and starch-iodine point to such infinite polyiodide chains in the latter as well.

Stable core-modified calixsmaragdyrins: synthesis, structure and specific sensing of the hydrogen sulfate ion.

Stable calixoxa- and calixthiasmaragdyrins containing three methine bridges and two direct bonds connecting the five pyrrole/heterocycle rings were synthesized by [3 + 2] condensation of dipyrromethane with 16-oxatripyrrane and 16-thiatripyrrane respectively under mild acid-catalyzed conditions. The compounds were characterized by HR-MS, 1D & 2D NMR, absorption and electrochemical techniques and the structure of calixoxasmaragdyrin was solved by X-ray crystallography. The crystal structure analysis indicated that the calixoxasmaragdyrin macrocycle was highly distorted due to the flexibility introduced by one sp(3)meso-carbon. The compounds show ill-defined absorption bands and irreversible oxidation and reduction waves which were attributed to the disruption of conjugation of the macrocycle by incorporation of one sp(3)meso-carbon. The anion binding studies indicated that the calixoxasmaragdyrin exhibited specific sensing ability for the HSO4(-) ion over other anions whereas calixthiasmaragdyrins did not even show an ability to bind anions.

Boron-dipyrromethene based reversible and reusable selective chemosensor for fluoride detection.

We synthesized benzimidazole substituted boron-dipyrromethene 1 (BODIPY 1) by treating 3,5-diformyl BODIPY 2 with o-phenylenediamine under mild acid catalyzed conditions and characterized by using various spectroscopic techniques. The X-ray structure analysis revealed that the benzimidazole NH group is involved in intramolecular hydrogen bonding with fluoride atoms which resulted in a coplanar geometry between BODIPY and benzimidazole moiety. The presence of benzimidazole moiety at 3-position of BODIPY siginificantly altered the electronic properties, which is clearly evident in bathochromic shifts of absorption and fluorescence bands, improved quantum yields, increased lifetimes compared to BODIPY 2. The anion binding studies indicated that BODIPY 1 showed remarkable selectivity and specificity toward F(-) ion over other anions. Addition of F(-) ion to BODIPY 1 resulted in quenching of fluorescence accompanied by a visual detectable color change from fluorescent pink to nonfluorescent blue. The recognition mechanism is attributed to a fluoride-triggered disruption of the hydrogen bonding between BODIPY and benzimidazole moieties leading to (i) noncoplanar geometry between BODIPY and benzimidazole units and (ii) operation of photoinduced electron transfer (PET) from benzimidazole moiety to BODIPY unit causing quenching of fluorescence. Interestingly, when we titrated the nonfluorescent blue 1-F(-) solution with TFA resulted in a significant enhancement of fluorescence intensity (15-fold) because the PET quenching is prevented due to protonation of benzimidazole group. Furthermore, the reversibility and reusability of sensor 1 for the detection of F(-) ion was tested for six cycles indicating the sensor 1 is stable and can be used in reversible manner.

Sensing Hg(II) in vitro and in vivo using a benzimidazole substituted BODIPY.

A multisignaling Hg(II) sensor based on a benzimidazole substituted BODIPY framework was designed, which displays excellent selectively toward Hg(II) in vitro and in vivo. Optical and fluorogenic measurements in solution reveal that the sensor can detect mercury ions at submicromolar concentrations, with high specificity. The detection of Hg(II) is associated with a blue-shift in optical spectra and a simultaneous increase in the fluorescence quantum yield of the sensor, which is attributed to a decrease in charge delocalization and inhibition of photoinduced electron transfer upon binding to Hg(II). Using several spectroscopic measurements, it is shown that the binding mechanism involves two sensor molecules, where lone pairs of the benzimidazole nitrogen coordinate to a single mercury ion. The utility of this BODIPY sensor to detect Hg(II) in vivo was demonstrated by fluorescence imaging and spectroscopy of labeled human breast adenocarcinoma cells. While average emission intensity of the sensor over a large number of cells increases with incubated mercury concentrations, spatially resolved fluorescence spectroscopy performed on individual cells reveals clear spectral blue-shifts from a subensemble of sensors, corroborating the detection of Hg(II). Interestingly, the emission spectra at various submicrometer locations within cells exhibited considerable inhomogeneity in the extent of blue-shift, which demonstrates the potential of this sensor to monitor the local (effective) concentration of mercury ions within various subcellular environments.

Synthesis of 3,5-bis(acrylaldehyde) boron-dipyrromethene and application in detection of cysteine and homocysteine in living cells.

Synthesis, characterization, and spectral and electrochemical properties of 3,5-bis(acrylaldehyde) BODIPY are described. The compound exhibited higher selectivity toward cysteine/homocysteine than toward other amino acids and thiol-containing compounds as shown by absorption and emission titration experiments and by experiments in living cells.

3,5-Diformyl-borondipyrromethene for selective detection of cyanide anion.

Cyanide is one of the most toxic inorganic anions, it is very harmful to human health but extremely useful in industrial activities. Herein, we used our recently reported boradiazaindacene (BODIPY) dye, 3,5-diformyl-borondipyrromethene (BODIPY 1) as an exclusive chemodosimetric and colorimetric sensor for CN(-) ion. Cyanide ion attacks the carbonyl groups of 1 via a nucleophilic addition reaction and converts to cyanohydrin which is reflected in the clear colour change as well as by the absorption, emission and electrochemical properties. Thus BODIPY 1 can be used as a colorimetric and chemodosimetric sensor for CN(-) ion. Furthermore, to show that the position of the formyl group on BODIPY plays an important role in the ability of BODIPY dye to act as a chemodosimetric sensor for CN(-) ion, we synthesized another formyl group containing BODIPY dye, 3, in which the formyl group is present at the para-position of the meso-phenyl group. (1)H NMR studies confirmed the formation of the cyanohydrin form of BODIPY dye 3 on addition of CN(-) ion but dye 3 cannot be used as a chemodosimetric sensor for CN(-) ion, as verified by absorption and fluorescence studies. The detection of cyanide with BODIPY dye 1 for biological application was also performed in MDA-MB-231 cells.

Synthesis, spectral, electrochemical, and anion binding properties of 3,5-bis(dipyrromethanyl) boron-dipyrromethenes.

Four new boron-dipyrromethenes (BODIPYs) containing dipyrromethanyl substituents at 3,5-positions, bis(3,5-dipyrromethanyl) BODIPYs 5-8, were synthesized by treating their corresponding 3,5-diformyl BODIPYs 1-4 with excess pyrrole under mild acid catalyzed reaction conditions. The compounds 5-8 are stable and freely soluble in common organic solvents. One-dimensional, two-dimensional NMR, high resolution mass spectrometry (HRMS), absorption, fluorescence, and electrochemical techniques were used to characterize the compounds. The spectral and electrochemical studies indicated that dipyrromethanyl groups at 3,5-positions of BODIPY are less electron deficient compared to formyl groups at the same positions. The anion binding studies indicated that bis(3,5-dipyrromethanyl) BODIPY compounds containing four pyrrole NH groups showed preferential binding with F(-) ion over other anions, as confirmed by using NMR, fluorescence, and electrochemical studies.

Fabrication of unipolar graphene field-effect transistors by modifying source and drain electrode interfaces with zinc porphyrin.

We report a unipolar operation in reduced graphene oxide (RGO) field-effect transistors (FETs) via modification of the source/drain (S/D) electrode interfaces with self-assembled monolayers (SAMs) of 5-(4-hydroxyphenyl)-10,15,20-tri-(p-tolyl) zinc(II) porphyrin (Zn(II)TTPOH) molecules. The dipolar Zn(II)TTPOH molecules at the RGO/platinum (Pt) S/D interface results in an increase of the electron injection barrier and a reduction of the hole-injection barrier. Using dipole measurements from Kelvin probe force microscopy and highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) calculations from cyclic voltammetry, the electron and hole injection barriers were calculated to be 2.2 and 0.11 eV, respectively, indicating a higher barrier for electrons, compared to that of holes. A reduced gate modulation in the electron accumulation regime in RGO devices with SAM shows that unipolar RGO FETs can be attained using a low-cost, solution-processable fabrication technique.

3,5-Diformylboron dipyrromethenes as fluorescent pH sensors.

A series of boron dipyrromethene (BODIPY) dyes containing two aldehyde functional groups at the 3 and 5 positions have been synthesized in low-to-decent yields in two steps. In the first step, the meso-aryl dipyrromethanes were treated with POCl(3) in N,N-dimethylformamide to afford 1,9-diformylated dipyrromethanes. In the second step, the diformylated dipyrromethanes were first in situ oxidized with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone and then reacted with BF(3)·OEt(2) to afford 3,5-diformylboron dipyrromethenes. The X-ray structural analysis indicated that the aldehyde groups are involved in intramolecular hydrogen bonding with fluoride atoms, which may be responsible for the stability of the diformylated BODIPY compounds. The presence of two formyl groups significantly alters the electronic properties, which is clearly evident in downfield shifts in the (1)H and (19)F NMR spectra, bathochromic shifts in the absorption and fluorescence spectra, better quantum yields, and increased lifetimes compared to 3,5-unsubstituted BODIPYs. Furthermore, 3,5-diformylboron dipyrromethenes are highly electron-deficient and undergo facile reductions compared to unsubstituted BODIPYs. These compounds exhibit pH-dependent on/off fluorescence and thus act as fluorescent pH sensors.