Laser-free Confocal Facility
Make
Aurox Ltd UK / Olympus
Model
Clarity High Speed (100 fps)/ Inverted Motorized Research Microscope- IX83
Facility Status
Working
Date of Installation
Facility Management Division
Centre for Sophisticated Research Facilities (CSIF- IoE Funded*)

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Category

  • Microscopy and Imaging » Confocal Microscopy

Booking Details

Booking available for
Internal and External Both
Available Equipment/ Mode of use
1. Confocal mode (Speed 100 fps) :
. Channels: DAPI, GFP, DsRed, CY5
. Z- stack imaging
. Real time live imaging
. Time-lapse imaging
. Modes Available: High Sectioning, Medium Sectioning and High Signal
2. Florescence microscope mode(Olympus IX83):
. Z- stack imaging
. Time-lapse imaging
. Multi-Colour imaging

Facility Management Team and Location

Faculty In Charge
Prof. Anirban Banerjee
abanerjee@iitb.ac.in
022-2576-7794
Facility Operator
Ms. Saranya Ajesh
saranya@iitb.ac.in
022-2159-6746
Facility Management Members
Prof. Anirban Banerjee (BSBE)
Prof. Shamik Sen (BSBE)
Prof. Rajesh Patkar (BSBE)
Prof. Shobhna Kapoor (Department of Chemistry)
Prof. Abhijit Majumder (Chemical Engineering Department)
Department
Biosciences and Bioengineering
Lab Email ID
saranya@iitb.ac.in
Facility Location
Imaging Facility Room, Ground floor, Department of Biosciences & Bioengineering, I.I.T Bombay, Powai, Mumbai - 400 076
Lab Phone No
022-2159-6746

Facility Features, Working Principle and Specifications

Facility Description

Facility Description

The High-throughput Imaging Facility provides a high-speed multi-channel imaging technique with significantly lower photobleaching than a conventional Point Scanning Laser Confocal Microscope.

This is a compact and fast confocal system that uses Aurox’s patented structured spinning disc and illumination technology to achieve high-resolution and high-quality confocal images. 

  • The resolution is about 300nm X 300nm in X-Y and 0.6um in Z direction. 
  • The laser-free confocal unit is coupled to an IX83 Olympus microscope.
  • On-stage incubation system with temperature and CO2 control for long-time live cell imaging.
  • Aurox spinning disk uses a grid pattern instead of pin holes for optical sectioning to achieve >50% light efficiency with the speed of a spinning disk.
Features Working Principle
  • Low photo-bleaching
  • Advanced image analysis software
  • Full 5D confocal imaging control.
  • Imaging across the scales with Clarity - 1000 microns to 0.2 microns
  • High compact and easy to use.

    Working Principle:- 

The unique optical system inside the Clarity LFC allows capturing of the images both transmitted (T) and reflected (R) by the disc to easily differentiate between in-focus+ out of Focus and out-of-focus information. Computer subtraction of these images (T–R) creates a sectioned image whereby all out-of-focus blur is effectively suppressed and only the sharp in-focus image of the sample is retained.

Using a spinning disk as the modulator/demodulator virtually eliminates residual imaging artifacts. This structured illumination pattern is used to both modulate the illumination field and demodulate the light emerging from the sample.

Body Specification

Aurox Confocal mode:

•Aurox patented structured illumination spinning disk

•NanoSRRF (Super Resolution Radial Fluctuations) Deconvolution

•Hamamtsu Fusion BT

•High end Xeon W-2245 core workstation

•sCMOS Hamamatsu Flash 4 V3 (95% QE) with Camera Link

•Cool LED pE 800 Light Source

Fluorescence microscope mode:

•Olympus IX83 with motorized X-Y-Z and Magnification of

    10X /0.3NA (air), 40X /0.60 NA (air), 60X /1.42NA (oil), 100X /1.45NA(oil)

Instructions for Registration, Sample Preparation, User Instructions and Precautionary Measures

Instructions for Registration

Internal users:

Users within IIT Bombay can apply online. Users are encouraged to be present during analysis by making a prior appointment. If a user cannot be present, a mail should be sent immediately to  saranya@iitb.ac.in along with the details of the sample as mentioned in the form

  • The slot is for 2 hours, the charges mentioned are per slot.
  • The experimental data provided is only for research/development purposes. These cannot be used as certificates in legal disputes.

External users:- 

Users can send an email to saranya@iitb.ac.in to apply for a slot.

Instruction for Sample Preparation

1. Seal the fixed samples between the glass slide and the cover slip.

2. When applying for a slot, please specify the fluorophores used in your sample, including their excitation and emission spectra.

3. Live samples should be in glass coverslip-bottom Petri dishes or 6, 12, 24, and 48 well plates.

4. Please use imaging Petri dishes with coverslip bottoms if oil immersion objectives are to be used.

Instructions for users: - 

  1. Standard slot durations are for 2 hours for fixed samples.
  2. USB drives are strictly prohibited to minimize virus-related issues. The data can be shared in the cloud. All data must be transferred within 7 days of imaging. Without exception.

Charges for Analytical Services in Different Categories

Applications

  • Live cell time lapse imaging
  • Fast dynamic imaging
  • Fixed cell imaging
  • Developmental Biology
  • Plant Biology
  • Stem cell research
  • Bioluminescent proteins

Sample Details

Gases allowed

CO2 

SOP, Lab Policies and Other Details

Publications

NA