Category
- Microscopy and Imaging » Optical Microscopy
- Spectroscopy and Spectrometry » Optical Spectroscopy
Booking Details
Facility Management Team and Location
Facility Features, Working Principle and Specifications
Features of TCSPC:
- Up to 1,000 measurements per second- ideal for kinetics studies
- monochromator wavelength selection
Measurement modes
- Lifetime – measure 60 ps to 1-2 microseconds
- Anisotropy – reconvolution analysis to resolve shorter rotational correlation times.
- Time-Resolved Emission Spectra (TRES) – Collect up to 100 wavelength dependent decays.
- Steady-State.
- Features of MicroTime 200 :
- Complete confocal system with laser combining unit, inverted microscope body, and dual-channel detection unit
- Turn-key diode lasers for multicolor excitation of 405, 440, 532 and 640 nm
- Two parallel detection channels using application-optimized detection with SPADs.
- Time-Correlated-Single Photon Counting (TCSPC) and TTTR mode for investigation of fast dynamics in FCS and FLIM
- Piezo scanning for 2D- and 3D-lifetime imaging and accurate point positioning
- Data acquisition, analysis and visualization software SymPhoTime 64
Principle of TCSPC:
Time-correlated single photon counting (TCSPC) is a common technique to measure fluorescence decays in the time domain. In principle, single photon events are detected, and their time of arrival is correlated to the laser pulse, which was used for excitation of the sample.
Principle of Fluorescence Lifetime Imaging Microscope (FLIM):
FLIM is a fluorescence imaging technique that resolves and displays the lifetimes of individual fluorophores rather than their emission spectra. The fluorescence lifetime is defined as the average time that a molecule remains in an excited state prior to returning to the ground state by emitting a photon.
Principle of Fluorescence Correlation Spectroscopy (FCS):
Fluorescence Correlation Spectroscopy (FCS) is a correlation analysis of temporal fluctuations of the fluorescence intensity. FCS enables the determination of important biochemical parameters such as the concentration and size or shape of the particle (molecule) or viscosity of their environment
- Internal users who have shown interest in getting trained in TCSPC and or in MicroTime 200 instruments, for this purpose, three-day training sessions are conducted to become an authorized users.
- Users can choose to get trained in any one or both the instruments according to the requirement of their lab.
- We give three days of training. Out of which, two days students are trained. On the third day, students can bring their own samples to have a hands-on session as they are encouraged to perform experiments under the guidance of staff operator, to become more familiar with the instrument.
- After becoming an independent operator, user shall also need to perform internal/external samples of other user requests. This will be a maximum of 4 hours per week (or even less) depending on the sample load.
Instructions for Registration, Sample Preparation, User Instructions, Precautionary Measures and Charges
Registration Process:
Internal Users:
Users within IIT Bombay can apply from http://drona.ircc.iitb.ac.in. The form should be completely filled up and all the sample details must be provided as indicated in the requisition form. Samples should be brought by the user at the allotted time and the user should be present at the time of analysis by taking prior appointments.
For TCSPC, maximum of 10 decays can be recorded per booking. The charges mentioned in the form are per decay recorded.
➢ For FLIM, the slot is of 2 hours and maximum 4 number of samples are allowed in a slot, the charges mentioned are for per slot.
External Users:
Since the registration portal is not accessible from outside IIT Bombay, External users must send a mail at the respective email-ids for registration. Students can be either present for the analysis or send via post at our facility address for sample analysis.
This below process is the same for both internal and external users. Following sample details should be mentioned. Users have to select the type of measurements:
- Time Correlated Single Photon Counting (TCSPC)
(No of decays, Excitation wavelengths ranges are from 275-315 nm and 365-510 nm, Emission wavelength, type of sample-solid/liquid)
- Fluorescence Lifetime Imaging Microscopy (FLIM)/ Fluorescence correlation Spectroscopy (FCS)
(No of samples, excitation wavelengths- 405,440,532,640 nm, Emission wavelength, type of sample-solid/liquid, description of the sample to be imaged, fluorophores present in the sample)
Appointment: The users will be informed about their date and timeslot by email.
Sample Submission: Samples are to be submitted at the time of registration or brought along on the date of their appointment for sample analysis.They can come in-person or can send their samples along with a letter from their Institute Original Letter Head for registration stating that the analysis is for research purposes. The letter should be addressed to Prof.Arindam Chowdhury (Incharge), Dept of Chemistry, IIT Bombay, Powai, Mumbai-400076
Users have to submit a receipt of payment along with the registration form and samples or else send through email also.
Results: Expected time for sending the result is at least three to four weeks after receiving the samples, depending on the sample load at the facility.
After the data acquisition is done the results will be sent by email for users.
For TCSPC:
For solid powder samples 10/15 mg of sample will do and for solution 10-50 μM of sample in your preferred solvent.
For MicroTime 200:
1)Solution samples should be well dispersed.
2)Samples for microscopy experiments should be sealed between glass slide and coverslip
3) For any further query, kindly contact on Email: trfs@iitb.ac.in
Contact: 022-2159-6895
1) For internal users, the designated lab operator who has received training for handling the instruments will be present for sample analysis.
2) Users are requested to bring all required samples, solvents, distilled water and washing acetone (if required), micropipettes and tips, cuvettes etc. For the analysis.
Applications
Application of TCSPC:
- Determination of Fluorescence lifetime.
Applications of MicroTime 200:
- Single Molecule Spectroscopy/Detection
- Fluorescence Lifetime Imaging (FLIM)
- Fluorescence Correlation Spectroscopy (FCS)
- Fluorescence Lifetime Correlation Spectroscopy (FLCS)
- Foerster Resonance Energy Transfer (FRET)
- Fluorescence Anisotropy (Polarization)
- Time-Resolved Photoluminescence (TRPL)
Sample Details
Not applicable
NO
Not applicable
Not applicable
NIL
Not applicable