Overview of Nikon
The Nikon A1 and C2 confocal microscopes systems can be easily expanded to Fluorescence Lifetime Imaging (FLIM) and Fluorescence Correlation Spectroscopy (FCS) capability using the ISS Upgrade Kits. External detectors are added to the system; photon counting acquisition ensures sensitivity to low signals. Data is acquired using one of two modalities: FastFLIM (digital frequency domain); or SWISS card (digital TCSPC, time-domain). While the choice depends upon the user preference, FastFLIM provides directly data in a suitable format for phasor plot analysis. The package includes VistaVision for data acquisition and processing.
The Nikon FCS and FLIM Upgrade Kit includes the following items:
|FastFLIM Unit or TCSPC Unit||Each unit accepts the output signals (via BNC) from the detectors and the synchronization signal from the Olympus confocal head.|
|Laser Launcher (not required for multiphoton systems)||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||High-performance computer, 32 GB RAM, with a 32" flat monitor|
Product Specifications for Nikon
Acquisition and Analysis Software
- VistaVision by ISS
Fluorescence Fluctuations Spectroscopy (FFS) Measurements
- Fluorescence Correlation Spectroscopy (auto- and cross-correlation)
- Photon Counting Histogram (PCH)
- FFS measurements at target XYZ locations in an image
- FLCS, Fluorescence Lifetime Correlation Spectroscopy
- Number & Brightness (N&B)
Imaging Module Measurements
- Single-point (intensity, polarization, lifetime)
- Single plan and z-stack (polarization images, Ratiometric, FLIM)
FLIM images (digital frequency-domain) (single plane and z-stack)
- Acquired in digital frequency-domain (DFD). The routine acquires simultaneously a FLIM image and a steady-state image.
FLIM images time-domain (single plane and z-stack)
- Acquired in time-correlated single photon counting (TCSPC)
Single Molecule Module
- Burst Analysis
- FRET and Correlation Methods
- PIE-FRET Methods
- Laser diodes: 370 nm - 1000 nm
- Single-photon pulsed lasers
- Multi-photon lasers
- Models for 3-, 4-, 6-laser. Light is delivered to the microscope through a single-mode fiber optic.
- GaAs PMT (Model H7422P)
- Hybrid PMTs (Model R10467U)
- Pixel, Line, Frame
FLIM Image Data Acquisition Minimum Dwell Time
- 6 µs/pixel
- High-performance Processor, 32 GB RAM, Windows 10, 64-bit
- 32" monitor, 2556 x 1440 resolution
“Photoluminescence enhancement and high accuracy patterning of lead halide perovskite single crystals by MeV ion beam irradiation.” Palei, M., Motapothula, M., Ray, A., Abdelhady, A.L., Lanzano, L., Prato, M., Panda, J.K., Scarpellini, A., Pellegrini, V., Primetzhofer, D., Petralanda, U., Manna, L. & Dang, Z. Journal of Materials Chemistry C, 8(29), pp. 9923–9930, 2020. doi: 10.1039/d0tc02326d.
“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. Journal of Biophotonics, 12(12), 2019, Aug. doi: 10.1002/jbio.201900164.