Gandhi Sivaraman

Here, we synthesized two thiol-based chemosensors for selective Hg ²⁺ detection; notably, Thiol-1 shows high selectivity even in the presence of competing metal ions.

Monitoring dynamic changes in mitochondrial pH in live cells using pH-sensitive fluorescent probe.

PH dynamically regulates diverse cellular functions and processes. At the Inner Mitochondrial Membrane (IMM), nanoscale pH gradients generated by the Electron Transport Chain (ETC) play a critical role in contributing to mitochondrial membrane potential that drives ATP synthesis and thermogenesis. However, tools to decouple pH gradients from the overall IMM potential in living cells are limited. This study integrates a fluorescent “benzo-indole” chromophore with a pH-sensitive “phenol” moiety into a single covalent skeleton to build a sensitive, red-shifted, cell-permeable pH probe (Mito-pH2). Mito-pH2 localizes inside mitochondria with high specificity presumably to the mitochondrial inner membrane by virtue of being an amphiphilic cation and can report dynamic changes in mitochondrial pH in living cells. Our design ensures that Mito-pH2 exhibits pH-sensitive dual-excitation and dual-emission peaks enabling ratiometric pH-sensing. Further, Mito-pH2 reports an increase in pH in the pH range of 3-9 through a striking colour change from yellow to purple making it a sensitive all-purpose colorimetric pH probe. A combination of DFT calculations and spectroscopy shed light on likely sensing mechanisms including photophysics. Quantitative live-cell fluorescence imaging reveals that Mito-pH2 can detect dynamic changes in mitochondrial pH upon extracellular pH modulation with little or no measurable cytotoxicity during live imaging. Red-emitting Mito-pH2 opens new avenues of quantitative mapping of physiological mitochondrial membrane pH and significantly enhances the repertoire of environment-sensitive and low-toxicity mitochondrial probes that link mitochondrial state and micro-environment.

Herein, we report a transition metal-free approach for the regioselective functionalisation of benzylic C(sp³)–H bonds using alcohols and carboxylic acids as the nucleophiles. Late stage functionalization has been done with complex drug molecules.

In this work, we report the development of nitrogen-doped carbon dots (NDCDs) as a drug carrier using quercetin (QC) as a model drug for anti-cancer drug delivery application. NDCDs were prepared by a simple hydrothermal method using Luffa acutangula as a carbon source. The characterization of QC-NDCDs was done by UV-vis spectroscopy, fluorescence spectroscopy, zeta potential measurements, high-resolution transmission electron microscopy (HR-TEM), Fourier transform infrared (FT-IR) spectroscopy, X-ray diffraction (XRD), and Raman spectroscopy. The as-synthesized NDCDs have a small particle size with hydroxyl and nitrogen-containing groups (pyridinic and amide groups), enhancing the fluorescence properties, and were obtained in a good quantum yield (14%). Furthermore, the in vitro alamarBlue® assay revealed that the NDCDs-QC conjugate was nontoxic to colon cancer cells. This NDCDs-QC conjugate is able to kill cancer cells in the NDCDs-QC form compared to free QC as confirmed by in vitro MTT assay results. Thus, the developed NDCDs conjugate can be used as a promising drug delivery and bio-imaging vehicle in cancer therapy.

The first discovery of aggregation-induced emission (AIE), whereby luminogen aggregation plays a positive role in enhancing the light-emission efficiency, has piqued the interest of many researchers as it opens up a new avenue for the exploration of practically beneficial luminescent materials. Diverse AIE-active luminogens (or AIEgens) with tunable emission colours and very high quantum yields (up to unity) in the solid state have been extensively utilised in a broad range of fields including optoelectronics, energy and bioscience. In this article, we describe novel fluorene-based fluorogens that exhibit bright emission in the solid-state, mechanical stimuli-responsive optical properties and aggregation-induced emissive ability, and were able to modulate their donor and acceptor properties. The target compounds were synthesized by a Knoevenagel condensation followed by Suzuki cross-coupling reaction, which tends to result in good yields. The target cyanostilbenes (4a–4d) show different reversibly switched states with high contrast through morphology modulation and demonstrate solvatochromic, vapochromic, and AIE properties. These results strongly suggest that compound 4d has better properties than the other derivatives (4a–c) due to the presence of extended donor–acceptor ability. Moreover, density-functional theory (DFT) calculations strongly support the UV-Vis and fluorescence spectral studies. The formation of nano-flakes and cuboid-shaped nanocrystals was further confirmed by FE-SEM and AFM studies. The synthesized compound 4d displayed very bright emission in the solid state and in the aggregate state as compared with the other derivatives (4a–4c). These results might be due to the presence of high-color contrast, which is an advantage for elucidation and overcomes the challenges exhibited in live-cell imaging applications. Moreover, an MTT assay on live A549 cells incubated with the target compound (4d) showed very low cytotoxicity even at high concentrations.

Environment-sensitive molecular probes offer the potential for a comprehensive mapping of the complex cellular milieu. We present here a radically new strategy of multiplexing highly sensitive, spectrally tuned fluorescent dyes for sensing cellular microenvironment. To achieve this multicolor, ratiometric cellular imaging, we first developed a series of highly sensitive, tunable molecular rotors for mitochondrial imaging, with emission wavelengths spanning the visible spectrum. These fluorogenic merocyanine dyes are all sensitive to solvent viscosity despite distinctive photophysical features. Our results show that merocyanine dyes can show a rotor-like behavior despite significant changes to the conventional donor–acceptor or push–pull scaffolds, thereby revealing conserved features of rotor dye chemistry. Developing closely related but spectrally separated dyes that have distinct response functions allows us to do ″two-color, two-dye″ imaging of the mitochondrial microenvironment. Our results with multidye, combinatorial imaging provide a direct visualization of the intrinsic heterogeneity of the mitochondrial microenvironment. The overall mitochondrial microenvironment (including contributions from local membrane order) as reported through two-color fluorescence ″ratio″ changes of multiplexed rotor dyes shows dynamic heterogeneity with distinct spatiotemporal signatures that evolve over time and respond to chemical perturbations. Our results offer a powerful illustration of how multiplexed dye imaging allows the quantitative imaging of mitochondrial membrane order and cellular microenvironment.