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Nikon, FCS and FLIM
The Nikon A1 and C2 confocal microscopes series can be easily upgraded to Fluorescence Lifetime Imaging (FLIM) and Fluorescence Correlation Spectroscopy (FCS) capability using photon counting detection. Data are acquired using one of two modalities:
- the FastFLIM (digital frequency domain); or
- the TCSPC (time-domain).
FastFLIM (proprietary technology) is the new digital frequency domain (DFD) approach to FLIM measurements. Its advantages, when compared to the TCSPC are twofold:
a) the short time required for data acquisition; and
b) the higher sensitivity of the technique (due to the 100% duty-cycle).
Alternatively, FLIM data can be acquired using the TCSPC method, or both methodologies can be implemented on the same instrument.
The Nikon, FCS and FLIM upgrade package includes the following items:
|FastFLIM Unit or TCSPC Unit||It accepts the output (via BNC) from up to four PMTs of the confocal unit. The synchronization signal from the Nikon confocal head is connected to the unit.|
|Detectors||2-detectors coupled to the descanned port confocal head of the LSM microscope; or to the non-descanned port (for multiphoton instruments)|
|Laser Launcher||Available for 3-, 4- and 6-lasers. The lasers beams are superimposed and the output of the laser launcher is connected to the microscope by using a fiber optic.|
|Computer Running VistaVision by ISS||A separate computer, with a 27" flat monitor|
|Acquisition and Analysis Software||
|Fluorescence Fluctuations Spectroscopy (FFS) Measurements||
|Imaging Module Measurements||
|FLIM images (digital frequency-domain) (single plane and z-stack)||
|FLIM images time-domain (single plane and z-stack)||
|Single Molecule Module||
|FLIM Image Data Acquisition Minimum Dwell Time||
Figure 1. Schematics of the upgrade package for the Nikon Frequency-Domain Confocal Microscope.
Figure 2. Schematics of the upgrade package for the Nikon Time-Domain Confocal Microscope.
Below is a list of selected publications featuring the Nikon, FCS and FLIM upgrade from ISS.
Photoluminescence Enhancement and High Accuracy Patterning of Lead Halide Perovskite Single Crystals by MeV Ion Beam Irradiation
Palei, M., Motapothulab, M, Ray A., Abdelhadya, A.L., Lanzanod, L., Prato, M., Panda, J.K., Scarpellini, A., Pellegrinif, V., Primetzhofer, D., Petralanda, U., Manna, L., Dang, Z.
J. Mater. Chem. C, 2020,8, 9923-9930
Chromatin nanoscale compaction in live cells visualized by acceptor-to-donor ratio corrected Förster resonance energy transfer between DNA dyes.
Pelicci, S., Diaspro, A., Lanzanò, L.
J Biophotonics. 2019 Jul 31:e201900164. doi: 10.1002/jbio.201900164. [Epub ahead of print]