National Institute of Plant Genome Research
Digital India   Azadi Ka Amrit Mahotsav     
 
SAHAJ-Platform for Measurement of Free Radicals
Overview
Free radicals are highly reactive chemical species generated from metabolic activity of cell under both normal and stress condition which have pivotal role in many biological processes. These free radical species play important role in signal transduction, regulation of gene expression and numerous cellular physiologies. Thus accurate quantification of these free radicals can provide insightful inferences to researchers regarding the system under study.

The Platform for measurement of Free Radicals (PMFR) at National Institute of Plant Genome Research (NIPGR) funded by Department of Bio-technology (DBT) under Scientific Infrastructure Access for Harnessing Academia University Research Joint Collaboration (DBT-SAHAJ) is one-of-its kind national platform providing state of the art advanced instruments for free radical research in both plant and animal systems. The facility provides high precision instruments for study of respiration, Internal oxygen flux dynamics (invasive and non-invasive), nitric oxide quantification, fluorescent detection of free radicals etc.
Instruments
Following equipments are available in the PMFR facility:
  1. nCLD CraNOx II (ecoPhysics, Switzerland)

  2. VisiSens 2D Oxygen imager with oxygen sensor RPSu4 (PreSens, Germany)

  3. VisiSens TD basic oxygen imaging system

  4. VisiSens TD Big area oxygen imaging system

  5. Oxygraph+ respiration measurement system (Hansatech, England)

  6. 4- Channel optic O2 transmitter- Optical fiber type (PreSens, Germany)

  7. 4- Channel optic O2 transmitter trace- Needle type (PreSens, Germany)

  8. Fluorescent microscope (Nikon Eclipse Ni, Japan)

  9. Incubator shaker

  10. Plant Growth Chamber

  11. Tissue homogenizer

  12. C- chamber gas mixing device

  13. Ultra deep freezer

1. nCLD CraNOx II

The nCLD 899 CY analyzer is the next generation instrument in high precision nitric oxide and NH3 measurement. Unique in speed and reliability, the nCLD 899 CY has modular designed and capable of detecting lowest quantities of NO, NO2 , NOx , NOx-amines and NH3 in the range of parts per trillion. The CraNOx II system is capable of simultaneously measuring NO, NOX , NO2 and O3. Furthermore, it can be expanded to assess NOY and NH3 concentrations.
 
 
2. Fluorescent microscope (Nikon Eclipse Ni)

The ECLIPSE Ni series is Nikon's flagship upright microscope, boasting great optical performance and high system expandability. Combined with high-performance objectives, it can accurately visualize microstructure of the sample with high resolution. Adopting a stratum structure that allows simultaneous mounting of multiple fluorescence attachments, it provides great flexibility to freely combine various accessories as needed. The fully motorized model enables automatic observation, responds to the needs of advanced research in which control of devices such as cameras and confocal systems is linked with microscope control, and supports a wide range of fluorescent ROS and RNS measurements in vivo.
3. VisiSens 2D oxygen imaging system
The VisiSens A1 is an imaging technology allowing easy 2D visualization of oxygen distributions in e. g. living, heterogeneous samples. The handheld digital camera records pixel by pixel sensor responses, capturing information of a whole array of sensor points. With VisiSens detector and oxygen sensor spatial and temporal changes of oxygen can be monitored. The SF-RPSu4 sensor foil for measuring oxygen allows non-invasive mapping of metabolic activities as well as changes over time periods from seconds to months. The fluorescent sensor foil is attached on a living or dead sample surface or a transparent glass or disposable vessel. A sensor film on the foil translates the oxygen content into a light signal. Read-out is done contactless with the imaging Detector Unit to acquire single images or automatically recorded time series.
 
 
4. VisiSens TD Basic oxygen imaging system

The VisiSens TD images large areas and gathers information on spatial and temporal analyte changes, e. g. in large scale pH and CO2 imaging of root-soil interactions within rhizotrons or when analyzing biogeochemical processes in sediments. It is also well suited for application in sensitive processes, e. g. metabolic activity of living samples or tissue engineering, where O2 gradients over a cross section of a 3D graft can be monitored.
5. VisiSens TD Big Area oxygen imaging system

The VisiSens TD images large areas and gathers information on spatial and temporal analyte changes, e. g. in large scale pH and CO2 imaging of root-soil interactions within rhizotrons or when analyzing biogeochemical processes in sediments. It is also well suited for application in sensitive processes, e. g. metabolic activity of living samples or tissue engineering, where O2 gradients over a cross section of a 3D graft can be monitored.
 
 
6. VisiSens TD Big Area oxygen imaging system

The Oxygraph+ oxygen electrode system provides PC control of oxygen uptake or evolution measurements across a broad range of applications from studies of mitochondria and cellular respiration to measurements of isolated chloroplast suspensions in photosynthesis research applications with up to 100% oxygen concentration. The Oxygraph+ system consists of a highly sensitive S1 Clark-type polarographic oxygen electrode disc mounted within a DW1/AD electrode chamber and connected to the Oxygraph+ electrode control unit. The DW1/AD provides a highly versatile solution to measurements of dissolved oxygen in liquid-phase samples of between 0.2ml - 2.5ml with its clear cast acrylic construction providing excellent sample visibility & uniform illumination. Precise temperature control of the sample and electrode disc can be achieved by connecting the water jacket of the DW1/AD to a thermoregulated circulating water bath.
7.  4- Channel optic O2 transmitter- Optical fiber type

This multi-channel oxygen meter is ideally suited for benchtop applications. It is compatible with non-invasive sensors of type PSt3 (detection limit 15 ppb dissolved oxygen, 0 - 100 % oxygen). Each channel of OXY-4 SMA has separate temperature compensation, so most precise measurements in environments with changing temperatures can be taken. A polymer optical fiber (POF) is needed to transfer excitation light to the sensor and the sensor response back to the meter. POF enables non-invasive and non-destructive measurements to be made from the outside through the wall of a transparent or slightly colored container.
 
 
8. 4- Channel optic O2 transmitter- Needle fiber type

This sensor is a miniaturized optical oxygen sensor designed for micro-invasive measurement of oxygen compatible with 4-channel oxygen meter. It is mounted inside a stainless steel needle, which protects the optical fiber and oxygen-sensitive sensor tip. This design is optimal for easy penetration of tissue

Contact Us:

PI
Dr. Jagadis Gupta Kapuganti

CO - PI
Dr. Swarup Parida
Dr. Sabhyata Bhatia

External users are requested to contact the facility - pmfrsahaj@gmail.com

Sample preparation protocols and training will be provided by the facility
Links
  1. Price List

  2. Sample Submission form

  3. Internal Users

  4. External Users

  5. Workshop and Training

Staff
  1. Dr. Priyanka Babuta (Project Scientist I)

  2. Dr. Deepak Saini (Senior Project Associate)