A. Surgical Equipment:
Various animal models have been developed in our lab to study the mechanism of degenerative disorders related to eye and brain. Strategies such as stem cell transplantation, screening of herbal extracts etc are also being employed to develop therapeutics. To carry out these tasks our lab is equipped with various sophisticated instruments that aid cell & molecular analysis, electrophysiological examination and microsurgeries.
1. LEICA SD6 MICROSCOPE WITH EC3 CAMERA
Leica Microsystems
First stereomicroscope in market with completely apochromatic optics, corrects perfectly chromatic aberrations, eliminates interfering color fringes and renders even the finest detail, ultra sharp. Contrast, brilliance, sharpness, resolution, color fidelity and reproduction accuracy are unsurpassed. It is used in lab to carry out various surgical procedures and animal models of retinal and brain injury such as Mid cerebral artery occlusion (MCAO), Pterygopalatine artery occlusion (PPA), NMDA have been established in the lab.
2. STEREOTAXIS APPARATUS
Stoelting
The Stoelting Just For Mouse™ is ideal for researchers in need of a versatile, reliable instrument for stereotaxic procedures with mice and other small rodents. Precision alignment when using the Just For Mouse ensures accurate placement of electrodes, micropipettes, cannula, and other devices. Stoelting’s Just For Mouse is the only instrument built especially for mice, and offers several advantages over competing “made for rat” instruments.
The small size, one of the smallest on the market, is ideal for laboratories with small spaces or for use in surgical suites that require transportation of equipment to and from surgical facilities. A zeroing function allows the user to target specific coordinates in the brain for injection, implantation, etc. Calculation of distance measurements is thus no longer necessary saving valuable time and improving accuracy. Ear bar posts may be independently adjusted in height—a unique feature of this stereotaxic instrument—to level the skull. Ear bars are composed of light Delrin material with tapered points or non-invasive rubber pads or jaw holder cuffs. Laser engraved scales show the vertical positions of the ear bars. Model organisms with soft skulls, such as the mouse and the neonatal rat, are difficult to secure firmly in a traditional stereotaxic instrument. The stainless steel ear bars used for adult rats are too heavy to provide a secure hold without compressing the sides of the skull inward (which closes the airways and impedes blood flow).
3. LASER PHOTOCOAGULATOR
Iridex
Photocoagulator is a fully integrated pattern scan laser system designed to injure or treat retina using a single spot or a predetermined pattern array of up to 56 spots. It is used to perform laser coagulation surgery on mice, laser spots are used to finely cauterize ocular blood vessels to create a Glaucoma model.
B. Analytical Technologies:
Analysing the results at histological, biochemical and molecular levels is an indispensable part of any research. Only result analysis can validate the procedures involved in an experiment. Lab is well furnished with the latest technologies involved in analysis of results at various levels. These technologies include conventional Thermal cycler to Real Time PCR; ELISA; UV Spectrophotometer; Cryostat; Fluorescent Microscope; Inverted Microscope; Refrigerated centrifuge; morris water maze.
1. REAL TIME PCR
Applied Biosystems
The StepOne system uses fluorescent-based polymerase chain reaction (PCR) reagents to provide quantitative detection of target nucleic acid sequences (targets) using real-time analysis, qualitative detection of targets using post-PCR (endpoint) analysis, qualitative analysis of the PCR product (achieved by melt curve analysis that occurs post-PCR).
The instrument has a 48 well platform and features a three-color filter set that supports the following Applied Biosystems dyes: FAM dye, JOE dye, ROX dye, VIC dye, SYBR Green dye.
It is used to amplify and simultaneously quantify a targeted DNA molecule, to study the genetic expression of particular gene over time, such as response of tissue and cell culture to an administration of a pharmacological agent etc.
PCR THERMAL CYCLER
Biorad
Conventional thermal cycler offers a comprehensive set of convenient features in a small footprint. This compact thermal cycler features an intuitive touch-screen user interface to make running PCR easy. Thermal gradient technology allows you to quickly optimize your reaction in a single run. Self-adjusting heated lid that accommodates standard 0.2 ml thin walled pcr tubes, strips and plates. Thermal gradient technology permits simultaneous incubation at 8 different temperatures for optimizing reactions in single run. It is used in nucleic acid amplification, gene expression analysis, mutational analysis and is useful in both research and diagnostics.
2. UV SPECTROPHOTOMETER
Beckmancoulter
The DU Series 700 spectrophotometers with a wavelength Range from 190 to 1100 nm are advanced and affordable systems that generate accurate and reproducible measurements. A wide range of applications software is included and makes the DU Series 700 the perfect solution for life science, research and analytical labs. The narrow beam width of the system ensures excellent results for small samples in semi-microcells. With the focused beam, there is no need for masking, and the entire energy is utilized to provide a wide dynamic range and excellent linearity. Its main use in the lab is to quantify the DNA or RNA sample.
3. ELISA READER
Biorad
The iMark™ microplate absorbance reader offers reading speeds of 6 sec at single wavelength detection, onboard software capabilities including protocols for kinetic assays, a preprogrammed validation protocol, a built-in plate shaker, and a built-in thermal printer. The instrument can read flat-, U-, or V-bottom microplates or 8- or 12-well strip plates. The iMark reader can be used as a stand-alone instrument or controlled by a PC or Mac with Microplate Manager® 6 software. It is provided with six filters (415–750 nm). It is used for estimating various cytokine and growth factors in human and rodent samples.
4. REFRIGERATED CENTRIFUGE
Thermofisher Scientific
Thermo Scientific centrifuges deliver speed and flexibility for reliable separations—all in a compact footprint. Refrigerated centrifuge featuring refrigeration and advanced intelligent technologies that eliminate imbalances, simplify operation and ensure reproducible results. It is used in the lab for separation of various fractions of biofluids and nucleic acids.
C. Immunohistochemical analysis:
1. CRYOSTAT
Leica
cryostat
The Leica CM1510 S is the best-value cryostat available for all cryo- and critical applications such as Mohs surgery. The CM1510 S allows rapid freezing and sectioning of skin cancer in multiple thin layers. Time is crucial when sectioning frozen specimens – An actively cooled quick freezing shelf with defrost function and a powerful refrigeration system are provided to get rapid results. In a minimum of time, the specimen can be clamped and oriented with the easy-to-use orientation system. The cryochamber can be quickly and effectively disinfected at any time with ultra-violet light (UVC) for increased operator safety. UV surface disinfection significantly reduces the danger of infection from bacteria, spores and viruses. Cryostat is used in lab to obtain section of eye with the thickness upto 4 µm.
2. FLUORESCENT MICROSCOPE
Leica
The ergonomically designed Leica DM1000 with adjustable features and easy-to-use controls adapts to every user to improve comfort and convenience while working at the microscope. Its advanced capabilities, including fluorescence, make it well suited also for complex tasks. With its image acquisition, processing and analysis capabilities, and complete set of tools for automation, Leica Florescent microscope opens the door for new insights into cellular function. Its flexibility and versatility makes it a powerful system for multi-dimensional image acquisition and 3D reconstruction, and for measurements such as morphometry, colocalization and motility. Immunohistochemical analysis of retinal sections is done in lab using fluorescent microscope.
D. Behavioural Technologies:
1.MORRIS WATER MAZE
Stoelting
The Digital video tracking Morris water maze is a behavioural procedure designed to test spatial memory. The maze has advantages over conventional mazes such as the elimination of local cues, like scent traces, and the fact that there is no fixed escape formula. For the study of behavioural analysis and memory loss studies morris water maze experiment is employed in lab.
2. ROTA ROD
Almicro Instruments
Some of the key features of rota rod are that no adjustments required on sensing platforms, Digitally controlled, Five (5) lanes standard, One (1) to Five (5) lanes may be used in a test, adjustable test length, adjustable start speed, adjustable ramp speed, forward and reverse rotation mode, distance recorded in meters, Mice use 1¼ inch diameter drums, rats use 3¾ inch diameter drums (both supplied). In lab it is used to observe the behavior of animals after various surgical procedures.
E. INVITRO TECHNOLOGIES:
Besides in vivo experiments, in vitro analysis of cell lines is another specialty of the lab. The high demand for aseptic conditions in culture work is provided by the Laminar hood in which culture is maintained in sterile conditions.
1. LAMINAR HOOD
Sonar
The laminar flow hood provides an aseptic work area while allowing the containment of infectious splashes or aerosols generated by many microbiological procedures. In the lab it is used to perform in-vitro experiments involving cell culture. Besides this all other experiments like DNA extraction, RNA extraction all are carried out in another laminar hood dedicated for this purpose.
2. SONICATOR
Sartorius stedim biotech
The Labsonic is a convenient, handheld instrument that allows fast sonication with a maximum of 100 W output. Using a flow cell, even larger samples can be processed. Sonotrodes are made of titanium and are highly resistant to caviation forces, but they still show some wear with prolonged use. The length of sonotrodes is therefore determined automatically by the processor and the frequency adjusted to re-establish optimum output conditions. This also is a unique feature of LABSONIC® instruments and enables longer usage of the sonotrodes. It is used generally to disrupt cell membranes and release cellular contents, to fragment molecules of DNA, to remove dissolved gases from liquids by sonicating the liquid while it is under vacuum and to loosen the particles adhering to surfaces.
3. INVERTED MICROSCOPE
Nikon
NIKON inverted microscopes are industry leaders; renowned for optical quality, flexibility, modularity, ease of use, ergonomics, and mechanical precision. Each is highly versatile and capable of producing brighter, higher contrast, aberration-free images. It is used in the lab to examine growing cells.
F. Electrophysiological Equipment:
1. LASER DOPPLER:
Moor Instruments
The moorVMS-LDF™ laser Doppler blood flow and temperature monitor is a high performance, medical grade module for clinic or laboratory. Use of DSP technology brings you a portable, lightweight module featuring uncompromised specification, quality and reliability at a breakthrough price for more demanding research applications, moorVMS-PC supports simultaneous protocol control of pressure cuff, iontophoresis and skin heating modules for reproducible blood flow provocation as well data acquisition with the NEW moorVMS-DAQ 8 channel data acquisition module. In lab it is used to confirm that occlusion of arteries has occur and blood flow is halted.
2. ELECTRORETINOGRAM:
iWorx 214 Data Recorder with mouse ERG electrodes
This rugged, reliable, four-channel data recording unit is engineered to take years of classroom use and abuse. The device enclosure is constructed from extruded aluminum, with a minimum of student-accessible controls and adjustments. For most applications, setup is automatic and transparent to the user. With one mouse click, the unit can be configured for EEG, EMG (EOG) or ECG. The iWorx 214 includes a dual-channel biopotential amplifier designed for use on human subjects. Two additional channels accept practically any sensor you are likely to encounter, as well as voltage output of pH meters, spectrophotometers and other laboratory devices. The iWorx 214 also includes a fully functional built-in stimulator useful for a variety of human and animal labs. The recorder interfaces to the PC via the USB bus, which provides aggregate sampling at 100 kHz continuous, more than fast enough for just about any teaching experiment. The mouse ERG electrodes are used to record the response stimuli from mouse eye in the lab.
G. Storage Facilities:
Ultra-Low Temperature (ULT) Lab Freezers providing temperature upto -80oC and yielding a highly efficient refrigeration system are used to safely preserve samples over the long term. Besides, chemicals used in experiments; rodents and humans DNA, RNA samples, brain, eye sections etc are stored at the required temperature in other storage refrigeration system providing temperature of 40 C or 200 C.