Overview of FLIMsight PRO
The FLIMsight PRO is an all-in-one benchtop solution that seamlessly integrates 2-photon excitation, fluorescence lifetime imaging, and phasor plot analysis for real-time, label-free diagnostics. Its compact design and user-friendly workflow simplify advanced imaging for researchers and clinicians alike. Optimized for detecting key metabolic coenzymes such as NADH and FAD, the FLIM system features dual-channel fluorescence lifetime acquisition, enabling precise differentiation of metabolic states in biological tissues. The system’s integrated two-photon laser, operating at 780 nm with a 40 MHz repetition rate, ensures deep tissue penetration while maintaining high spatial resolution. Detection is achieved through fast photomultiplier tubes (PMTs) with a 150 ps instrument response function (IRF), allowing for highly sensitive and accurate lifetime measurements. Utilizing FastFLIM technology, the system captures lifetime decay dynamics with exceptional speed, while advanced analysis tools, including multi-exponential fitting and phasor plot analysis, enable intuitive and quantitative data interpretation. With its streamlined workflow, compact design, and fully integrated hardware and software, the FLIMsight PRO simplifies advanced fluorescence imaging, making cutting-edge FLIM applications more accessible to both biomedical researchers and clinical professionals.
Key Features of FLIMsight PRO
All-in-One Compact
Benchtop Solution
Fast Image Acquisition
(Dwell Time: 0.2 µs)
High Dynamic Range
Fully Automated
Easy and Fast Analysis
Using the Phasor Plot
2-Photon Laser
3D Volume Rendering
An example of 3D volume rendering with FLIMsight PRO. A stack of z-sectioning images acquired from a pollen grain, using the two-photon laser excitation at the 780 nm wavelength.
Hematoxylin and Eosin-stained Tissue
Image acquired from a hematoxylin and eosin-stained tissue using the two-photon laser excitation at the 780 nm wavelength and a hybrid PMT photon counting detector. Both are capable of fluorescence lifetime imaging (FLIM). Together with ISS FastFLIM technology, it is straightforward to collect the fluorescent decay data at each image pixel. The time-resolved data is quantitatively unmixed by the ISS VistaVision phasor plots module producing three separate images which represent the fluorescence intensity contributed exclusively by distinct fluorescent species. The merged image is constructed utilizing false red, green and blue colors for shortest, intermediate and longest lifetime species, respectively.
Two-Component Lifetime Unmixing using the Phasor Plot
Using the phasor plots of the FLIM images acquired from NADH in live cells and solution which only contains the free form, multi-image phasor analysis (MiPA) derives the phasor coordinate of the bound form of the cellular NADH. MiPA computes the fractions of both free and bound NADH for each pixel in the cellular NADH image, producing a cellular map depicting the ratios of free-to-bound NADH. Given the free and bound NADH lifetimes, it also generates the cellular NADH average lifetime map.
Tissue Imaging
Autofluorescence tissue image: Excitation: 375 nm, Emission: 425 nm, Objective: 20X NA0.75
(courtesy of Dr. Aneesh Alex, GSK)
Product Specifications for FLIMsight PRO
Computer & Monitor
- High performance computer loaded with OS Windows 11 64 bit
- 64 GB RAM & 2 TB SSD
- 34", 3440 x 1440 resolution, flat color monitor
2-P Laser
- 780 nm, 40 MHz Rep Rate
Galvano-Mirrors
- Set of two mirrors for XY scanning
Detectors
- Two PMTs, IRF<150 ps, 280 - 620 nm
Acquisition
- FastFLIM (Digital Frequency Domain FLIM)
Automation Control
- USB port
Power Requirements
- Universal power input: 110 - 240 V, 50/60 Hz, 400 VAC
Dimensions (cm)
- 57 (L) x 39 (W) x 22 (H)
Weight (Kg)
- 28.5 (without microscope)
Scan Modes
- Single Spot
- 1D Line (X, Y or Z)
- 2D Image (XY, XZ, YZ)
- 3D Image Stack (XYZ)
- Each can be combined with time (t)
Scan Settings
- Pixels Number: 1 x 1 to 4096 x 4096
- Dwell Time: 0.5 μs to infinity
- Optical Zoom: 1 to 40
- Field Diameter: 18 mm
Data Acquisition by Photon Counting
- Photon Mode
- Time Tagged Mode
- Time Tagged Time Resolved Mode
Data Display and Analysis
- Image enhancement by thresholding, contrast adjustment & smoothing
- Image false coloring with more than 50 LUTs & overlay of multiple channels
- Image quantification by advanced mathematical operations
- FLIM data analysis includes both fitting (up to 5 components) & phasor plots
Image Storage Formats
- ISS proprietary formats with meta information
- Export to TIFF, GIF, JPEG, PNG, BMP, AVI, TXT
- Plots can be saved & exported to GIF, TIFF, JPEG, PNG, Bitmap & Metafile formats
Example Configuration for FLIMsight PRO
Product Software for FLIMsight PRO
VistaVision
VistaVision is a complete software package for confocal microscopy applications including instrument control, data acquisition and data processing. Easy to use, the software has been developed in modular components that can be activated when a specific instrument configuration is selected. The modules include:
- FLIM/PLIM Imaging
- FFS
- smFRET
- Particle Tracking
