Research Interest :
Human embryonic stem cell cultures: Culturing of mouse fibroblast cells (Feeder cells) which acts as support for the growth of human embryonic stem cells (hESCs), both induced and spontaneous differentiation of human embryonic stem cells into retinal pigment epithelial cells (RPE)
Maintenance of human embryonic stem cells using enzymatic passaging, non-enzymatic-manual passaging protocols.
Experience on Proton beam radiation therapy experimental design and analysis
Elecetrophysiology: Electroretinography (ERG) for the analysis of various inherited retinal degenerative diseases
Optical coherence tomography (OCT) is an established medical imaging technique that uses light to capture micrometer-resolution, three-dimensional images from within optical scattering media (e.g., biological tissue such as retina).
Cell culture methods : analysis of cellular proliferation assays using trypan blue exclusion assay, WST-1 & MTT assays
Cell death analysis: using Caspase-3 activity, Colorimetric In-cell ELISA, Propidium Iodide staining
Analytical Techniques in Protein Chemistry: SDS-PAGE, 2D-gel electrophoresis, Mass Spectrometry and PMF analysis, Western blotting.
Protein Purification Techniques: Gel filtration and C5-12 RP- chromatography, use of desalting columns using FPLC, Akta purifier.
Chromatography techniques: Separation of micro molecules using Reverse phase high performance liquid chromatography (RP-HPLC), Thin Layer chromatography (TLC).
Immunological Techniques: Enzyme linked immunosorbent assay (ELISA), Luminex technologies: for the analysis of various proinflammatory and anti-inflammatory cytokines associated with ocular deseases.
Analysis of various cytokines using Luminex multiplex technology
Molecular biology techniques: Isolation genomic DNA, amplification by PCR, Real Time PCR for gene expression analysis. Identification of single nucleotide polymorphisms (SNPs) by restriction fragment length polymorphism (RFLP), Single strand conformational polymorphism (SSCP), ARMS-PCR method and also by highly specific technique such as denaturing high performance liquid chromatography (DHPLC), DNA sequencing and sequence analysis. Use of secondary structure predictions of RNA using bioinformatics tools.
Experience in the analysing software packages for Protein& nucleotide sequence analysis and general scientific data analysis - Protein data bank, gene bank and Swiss Prot analysis, Gel analysis and Gel documentation systems.
Gene expression analysis by Real time PCR analysis.
Immunocytochemical analysis using various image software.
Expertise in handling various laboratory instruments as Thermal Cycler, Gel Documentation System, Spectrophotometer, Protean IEF cell, BioPhotometer, High performance liquid chromatography (HPLC), Fast protein liquid chromatography (FPLC), Akta Purifier, Gradient Ultracentrifuge, Sonicator, Homogenizer, Lyophilizers, Spectro Fluorimeter, ELISA Reader, Real-time PCR, Polyacrylamide Gel Electrophoresis, Genetic screening – DNA sequencing and data analysis, Western blotting and ELISA.
Electrophysiology : Electroretinography (ERG)
Electroretinography (ERG) measures the electrical responses of various cell types in the retina, including the photoreceptors (rods and cones), inner retinal cells (bipolar and amacrine cells), and the ganglion cells. Electrodes are usually placed on the cornea and the skin near the eye, although it is possible to record the ERG from skin electrodes. During a recording, the patient's eyes are exposed to standardized stimuli and the resulting signal is displayed showing the time course of the signal's amplitude (voltage). Signals are very small and typically are measured in microvolts or nanovolts. The ERG is composed of electrical potentials contributed by different cell types within the retina, and the stimulus conditions (flash or pattern stimulus, whether a background light is present, and the colors of the stimulus and background) can elicit a stronger response from certain components.