The research laboratory at the department of ophthalmology has modules for histology, molecular biology, cell culture, flow cytometry and microbead analysis. We have an outstanding collection of aqueous and vitreous samples from both normal and pathological eyes.

Histology facilities

We have human histological samples of different ocular pathologies and these samples are supplied from eye banks and from other institutions. Our lab has the expertise and is well equipped to perform immunohistochemistry experiments to identify the different protein expression and the results are analyzed using three channel and fluorescent microscopes (Olympus 1X51). Our investigators are experts with considerable experience in performing histological analysis including immunohistochemistry, confocal microscopy, electron microscopy and histochemical staining of both human and animal tissue.

Molecular biology facilities

We have very well equipped molecular biology facilities with gradient polymerase chain reaction cycler (PCR; Bio-rad) to amplify a specific gene from gene pool and western blot (Bio-rad) to visualize our protein of interest. PCR has a capability to do 8 different temperatures for 8 rows at one run and step-up, step-down and hot start PCR reactions. The amplified gene reaction products evaluated using either horizontal agarose gel electrophoresis unit or vertical polyacrylamide gel electrophoresis (PAGE) units whereas protein products were visualized using SDS-PAGE units. Further, the amplified gene products and immune blots can be quantified and analyzed using chemi-doc gel documentation system (Bio-rad).

Cytology

Our lab has a laminar flow hood under which we work on all cell work, in aseptic and contained environment, free of contamination. We have two CO2 incubators for preservation of cells and cell work. We also have a liquid nitrogen tank for storing our precious cells.

We quantify our cells using automated cell counter (Vi cell XR) which not only count our cells but also gives 50 camera images. This also tells our viable cell size in micron, circularity and the number of live and dead cell in each camera image and the average cell size and circularity for 50 camera images.

We use plate reader from Biotek synergy HT which is useful to measure luminescence, fluorescence and absorbance for functional, proliferative and cytotoxic experiments. We quantify reactive oxygen species concentration, DNA/RNA/protein concentration, protein interactions and cytokine levels using this plate reader. Instead of saying microplate reader we should say as ‘macro reader’ based on its usage.

Flow cytometry

We also have a Quanta flow cytometer by Beckman Coulter for cellular analysis. This will also helpful to quantify the number of cells undergoing apoptosis and necrosis based on its specific fluorescent agent bound to cell. We use this instrument to identify the cells undergoing proliferative phase, resting phase in general cell cycle analysis in order to determine which phase cell cycle is affected by our treatment agent/condition.