Scientifica FLIM Upgrade Kit

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Scientifica FLIM Upgrade Kit

The Scientifica FLIM upgrade kit is an integrated and flexible Fluorescence Lifetime Imaging solution. When installed on a Scientifica multiphoton microscope, the FLIM upgrade enables simultaneous fluorescence intensity and fluorescence lifetime imaging, in up to two colour channels, using the versatile Picoquant TCSPC system.

By combining the high temporal resolution of intensity imaging with the detailed information on chemical environment provided by fluorescence lifetime imaging, highly quantitative data can be acquired.

Benefits

Flexible Integration

FLIM works with all Scientifica multiphoton scan heads and supports galvo and resonance imaging. Existing multiphoton systems are upgradeable to FLIM.

Simultaneous Intensity and Lifetime Imaging

Getting the best of both worlds: fast frame rates from regular fluorescence intensity imaging and the quantitative information from lifetime imaging.

Fast Photon-counting

FLIM at photon rates up to 1.5 Gcounts/second, turning the sample brightness into the limiting factor, rather than the detection system. Full support of rapidFLIM.

Dedicated FLIM Software

Quick access to new analysis routines. System runs ‘as usual’ for users not requiring FLIM. Proven technology.

Applications

FRET

• For the measurement of molecular interactions
• For quantification of biosensor measurements
• FLIM is the gold-standard for measuring FRET
• Straightforward access to FRET efficiency and fraction, with no need for acceptor photobleaching, or sets of multiple calibration images

Measurement of absolute ion concentrations

• Ratiometric calcium imaging using non-ratiometric indicators such as OGB-1
• Chloride measurements using quinine-based indicators such as  MQA or MEQ
• pH measurements

Alternative contrast

• Separate labelling with spectrally similar dyes
• Add meaningful contrast to (not just) autofluorescence imaging
• Contrast that is not dependent on dye concentration, but functional state

Comparison of maximum intensity and FLIM images

Images taken from the same autofluorescent plant leaf sample. Left: taken using FLIM. Right: taken using maximum intensity projection (MIP) imaging, The FLIM image shows areas that have the same brightness but very different lifetimes, which are not shown in the MIP image.
Two-photon excited autofluorescence of a plant leaf. Both images show a maximum intensity projection of the imaged volume. In the FLIM image (on the left) structures with very similar brightness can be clearly distinguished by their very different fluorescence lifetimes. In contrast, in the intensity image (on the right), these structures cannot be distinguished.
Zoomed in images of the same plant leaf autofluorescence. Left: Fluorescence lifetime image. Right: Intensity image
Zoomed in images of the autofluorescent plant leaf sample. Left: taken using FLIM. Right: taken using MIP imaging.

Design & Specifications

TCSPC Hardware
TCSPC Hardware
High time resolution pack: PicoQuant TimeHarp 260 Pico
Fast counting pack: PicoQuant MultiHarp 150
TCSPC time resolution and peak count rates
TCSPC time resolution and peak count rates
High time resolution pack: 25 ps, 40 MCounts/s
Fast counting pack: 80 ps, 1.5 GCounts/s
Software
Software
Intensity imaging: Scanimage 5/2016 or later
FLIM imaging: PicoQuant SymphoTime 64
Scanning modes
Scanning modes
Galvo: Mono- and bidirectional
Resonance: Only monodirectional
Detectors
Detectors
One or two hybrid detectors (PicoQuant PMA-Hybrid), each with an SMA output connector
Detector voltage control
Detector voltage control
Fixed, and set internally for optimal photon-counting efficiency
Intensity imaging detection
Intensity imaging detection
Analogue photon counting
Emission filters
Emission filters
25 mm Ø - one for each detector
Laser requirements
Laser requirements
Ultrafast pulsed laser, with a repetition rate of 80 MHz or less
Objective and Nosepiece Compatibility
Objective and Nosepiece Compatibility
Objectives: M32, M27, M25, RMS threaded objectives (<20mm back aperture)
Nosepieces: Ships with single objective changer with 2 locking sliders
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