LIFA vTau Camera for fluorescence lifetime imaging microscopy (FLIM)
LIFA vTau Camera for fluorescence lifetime imaging microscopy (FLIM)
Multi-LED
LIFA Software

LIFA vTau | Fluorescence Lifetime Imaging Microscopy

Modality

Fluorescence-Lifetime Imaging Microscopy

Configuration

Widefield, Confocal, TIRF, SPIM

Lifetime Resolution

10 ps

Lifetime Range

0.2 - 300 ns

FOV

420 x 420 µm (@ 20x, 512 x 512 pixels)

Speed

300 fps

LIFA vTau SPAD-powered FLIM Camera

LIFA vTau Fluorescence Lifetime Imaging Microscopy, in a nutshell

LIFA vTAU is a camera-based system for fast fluorescence lifetime imaging microscopy (FLIM), especially well-suited for live cell applications. Featuring vTAU, our versatile SPAD camera, the system allows for near instantaneous acquisition of lifetime images at unprecedented frame rates with high accuracy. The Lambert Instruments LIFA vTau is the fastest and easiest way to do fluorescence lifetime imaging microscopy (FLIM).

Available in versatile configurations dependent on your specific applications, the LIFA vTau system offers a turn-key solution for fluorescence lifetime imaging microscopy. Compatible with every fluorescence microscope with a camera output – including microscopes by Leica, Nikon, Olympus, and Zeiss; set up is easy and fast, with all hardware integrating seamlessly with our LIFA software, so you can focus on your experiment.

The advanced software instantly analyzes your data and presents the calculated fluorescence lifetimes visually. Recorded images are compatible with ImageJ, FIJI, MatLab and MetaMorph, while detailed statistical data can be exported to Excel worksheets.

LIFA vTau Fluorescence Lifetime Imaging Microscopy, in more details

Simplify experiments for researchers and imaging centers with the new vTAU SPAD camera; combining excellent light sensitivity with easy image acquisition and data analysis.

Minimize measurement duration, automate image acquisition, and simplify data analysis… factors of great importance for cell biology, cancer research and high-throughput screening.

LIFA vTAU easily integrates into any FLIM system, providing a plug-and-play experience that allows for switching between setups.

Key features & specifications

  • Complete solution: LIFA vTAU works with any brand of fluorescence microscope to form a fully integrated FLIM system for a complete solution from sample to data.
  • Multiple configurations: Suitable for widefield, spinning-disk confocal, light-sheet, and TIRF modalities, LIFA vTAU delivers an easy plug-and-play setup experience.
  • Fast, adaptive dynamic range: Utilizing the latest ultra-high sensitivity SPAD detector, featuring micro-lenses to improve light efficiency and using ultra-short exposures to optimize dynamic range, vTAU can capture up to 100 lifetime images per second in challenging light conditions.
  • Next level sensitivity: With the unique properties of SPAD, vTAU features excellent light sensitivity to minimize measurement duration, and enables noise-free readout of the detector.
  • Broad lifetime range: Matching a wide range of capabilities, vTAU operates from the sub microsecond down to picoseconds range.
  • Automatic data analysis: Dedicated LIFA software instantly calculates the fluorescence lifetime and presents it as a color-coded overlay on the original image.

LIFA system components

  • vTau camera: Versatile, plug-and-play FLIM camera for quick and easy setup using ultra-high sensitivity SPAD detector for up to 100 lifetime images per second.
  • Light source (Multi-LED / Multi-LASER): Excitation light sources for frequency-domain fluorescence lifetime imaging, supplied according to the wavelengths you require.
  • LIFA software: Seamless integration of all hardware for full system control, guiding through FLIM experiments from start to finish, it supports third-party hardware for a flexible and expandable system.

What is Fluorescence Lifetime Imaging Microscopy (FLIM) ?

Fluorescence Lifetime Imaging Microscopy (FLIM) is an advanced imaging technique used in microscopy to study the dynamic properties of fluorophores within biological samples. Unlike traditional fluorescence microscopy, which captures only the intensity of emitted fluorescence light, FLIM measures the time it takes for a fluorophore to return to its ground state after excitation. This information provides insights into the microenvironment of the fluorophores, allowing researchers to distinguish between different molecular species with overlapping fluorescence spectra and study molecular interactions in living cells. FLIM is particularly valuable in areas such as cell biology and medical research, enabling the visualization of subtle changes in cellular processes, the identification of biomolecular interactions, and the detection of cellular abnormalities at the molecular level. This technique has proven instrumental in advancing our understanding of complex biological phenomena and holds great potential for applications in diagnostics and drug development.

What about Time-Domain and Frequency-Domain FLIM ?

Time-domain and frequency-domain are two different approaches to perform Fluorescence Lifetime Imaging Microscopy (FLIM), each offering unique advantages in capturing temporal information about fluorescence decay.

Time-Domain FLIM:

    • Principle: In time-domain FLIM, the fluorescence decay of a fluorophore is directly measured in the time domain.
    • Procedure: A short-pulsed laser is used to excite the sample, and the resulting fluorescence emission is detected. The time it takes for the fluorescence intensity to decay from its peak to a fraction (often half) of its peak value is directly measured. This time is called the fluorescence lifetime.

Frequency-Domain FLIM:

  • Principle: In frequency-domain FLIM, the excitation light is modulated at a high frequency, and the phase shift and modulation of the resulting fluorescence signal are analyzed.
  • Procedure: A modulated excitation light source (LED or laser) is used to excite the sample. The fluorescence emission signal is demodulated to separate it from the excitation light. The phase shift and modulation are then used to calculate the fluorescence lifetime.

Frequency-domain FLIM doesn’t require pulsed laser nor advanced electronics which are often very expensive, and more complicated to use. Frequency-domain FLIM is also faster than time-domain FLIM. The LIFA vTau is a frequency-domain FLIM system, using a camera to measure in couple of frames the fluorescence lifetime simultaneously collected on each pixels.

LIFA vTau SPAD-powered FLIM Camera

LIFA vTau Fluorescence Lifetime Imaging Microscopy, in a nutshell

LIFA vTAU is a camera-based system for fast fluorescence lifetime imaging microscopy (FLIM), especially well-suited for live cell applications. Featuring vTAU, our versatile SPAD camera, the system allows for near instantaneous acquisition of lifetime images at unprecedented frame rates with high accuracy. The Lambert Instruments LIFA vTau is the fastest and easiest way to do fluorescence lifetime imaging microscopy (FLIM).

Available in versatile configurations dependent on your specific applications, the LIFA vTau system offers a turn-key solution for fluorescence lifetime imaging microscopy. Compatible with every fluorescence microscope with a camera output – including microscopes by Leica, Nikon, Olympus, and Zeiss; set up is easy and fast, with all hardware integrating seamlessly with our LIFA software, so you can focus on your experiment.

The advanced software instantly analyzes your data and presents the calculated fluorescence lifetimes visually. Recorded images are compatible with ImageJ, FIJI, MatLab and MetaMorph, while detailed statistical data can be exported to Excel worksheets.

 

LIFA vTau Fluorescence Lifetime Imaging Microscopy, in more details

Simplify experiments for researchers and imaging centers with the new vTAU SPAD camera; combining excellent light sensitivity with easy image acquisition and data analysis.

Minimize measurement duration, automate image acquisition, and simplify data analysis… factors of great importance for cell biology, cancer research and high-throughput screening.

LIFA vTAU easily integrates into any FLIM system, providing a plug-and-play experience that allows for switching between setups.

Key features & specifications

  • Complete solution: LIFA vTAU works with any brand of fluorescence microscope to form a fully integrated FLIM system for a complete solution from sample to data.
  • Multiple configurations: Suitable for widefield, spinning-disk confocal, light-sheet, and TIRF modalities, LIFA vTAU delivers an easy plug-and-play setup experience.
  • Fast, adaptive dynamic range: Utilizing the latest ultra-high sensitivity SPAD detector, featuring micro-lenses to improve light efficiency and using ultra-short exposures to optimize dynamic range, vTAU can capture up to 100 lifetime images per second in challenging light conditions.
  • Next level sensitivity: With the unique properties of SPAD, vTAU features excellent light sensitivity to minimize measurement duration, and enables noise-free readout of the detector.
  • Broad lifetime range: Matching a wide range of capabilities, vTAU operates from the sub microsecond down to picoseconds range.
  • Automatic data analysis: Dedicated LIFA software instantly calculates the fluorescence lifetime and presents it as a color-coded overlay on the original image.

LIFA system components

  • vTau camera: Versatile, plug-and-play FLIM camera for quick and easy setup using ultra-high sensitivity SPAD detector for up to 100 lifetime images per second.
  • Light source (Multi-LED / Multi-LASER): Excitation light sources for frequency-domain fluorescence lifetime imaging, supplied according to the wavelengths you require.
  • LIFA software: Seamless integration of all hardware for full system control, guiding through FLIM experiments from start to finish, it supports third-party hardware for a flexible and expandable system.

What is Fluorescence Lifetime Imaging Microscopy (FLIM) ?

Fluorescence Lifetime Imaging Microscopy (FLIM) is an advanced imaging technique used in microscopy to study the dynamic properties of fluorophores within biological samples. Unlike traditional fluorescence microscopy, which captures only the intensity of emitted fluorescence light, FLIM measures the time it takes for a fluorophore to return to its ground state after excitation. This information provides insights into the microenvironment of the fluorophores, allowing researchers to distinguish between different molecular species with overlapping fluorescence spectra and study molecular interactions in living cells. FLIM is particularly valuable in areas such as cell biology and medical research, enabling the visualization of subtle changes in cellular processes, the identification of biomolecular interactions, and the detection of cellular abnormalities at the molecular level. This technique has proven instrumental in advancing our understanding of complex biological phenomena and holds great potential for applications in diagnostics and drug development.

 

What about Time-Domain and Frequency-Domain FLIM ?

Time-domain and frequency-domain are two different approaches to perform Fluorescence Lifetime Imaging Microscopy (FLIM), each offering unique advantages in capturing temporal information about fluorescence decay.

Time-Domain FLIM:

    • Principle: In time-domain FLIM, the fluorescence decay of a fluorophore is directly measured in the time domain.
    • Procedure: A short-pulsed laser is used to excite the sample, and the resulting fluorescence emission is detected. The time it takes for the fluorescence intensity to decay from its peak to a fraction (often half) of its peak value is directly measured. This time is called the fluorescence lifetime.

Frequency-Domain FLIM:

  • Principle: In frequency-domain FLIM, the excitation light is modulated at a high frequency, and the phase shift and modulation of the resulting fluorescence signal are analyzed.
  • Procedure: A modulated excitation light source (LED or laser) is used to excite the sample. The fluorescence emission signal is demodulated to separate it from the excitation light. The phase shift and modulation are then used to calculate the fluorescence lifetime.

Frequency-domain FLIM doesn’t require pulsed laser nor advanced electronics which are often very expensive, and more complicated to use. Frequency-domain FLIM is also faster than time-domain FLIM. The LIFA vTau is a frequency-domain FLIM system, using a camera to measure in couple of frames the fluorescence lifetime simultaneously collected on each pixels.

LIFA vTau Applications

Live-Cell Imaging

  • Track lifetimes with the built-in time-lapse feature
  • Observe small and rapid transients (~2.5 s periods) that could not be noticed using conventional FLIM
  • Allows simultaneous recording
  • Immune to artifacts caused by cellular movements and signal transients
  • Enable video-rate lifetime imaging with simple calibration

 

Bacteria Research

  • Frequency Domain FLIM to analyze FRET in single bacterial cells [3]

High Throughput Screening

  • Multi-position fluorescence lifetime imaging (FLIM) screening method for screening bright fluorescent proteins (University of Amsterdam)
  • In experiments where the fluorescence lifetime is an important parameter

Images acquired with LIFA FLIM systems

Stitched lifetime image of Hap 1 cells expressing EPAC-sensor that shows a lifetime increase when cAMP concentration increases after stimulation with isoproternol
Stitched lifetime image of Hap 1 cells expressing EPAC-sensor that shows a lifetime increase when cAMP concentration increases after stimulation with isoproternol. Courtesy of the Netherlands Cancer Institute.
B.subtilis cells showing different lifetimes. Top row: original lifetime image of FRETing, non-FRETing, and mixed cells. Bottom row: same cells as top row, but categorized and colorized based on average cell lifetime.
B.subtilis cells showing different lifetimes. Top row: original lifetime image of FRETing, non-FRETing, and mixed cells. Bottom row: same cells as top row, but categorized and colorized based on average cell lifetime. Courtesy of University of Groningen.
Top row: Fluorescence intensity images (colorized). Bottom row: Corresponding fluorescence lifetime images (colorized). The average fluorescence lifetime of the cells increases over time.
Top row: Fluorescence intensity images (colorized). Bottom row: Corresponding fluorescence lifetime images (colorized). The average fluorescence lifetime of the cells increases over time. Courtesy of the Netherlands Cancer Institute.
Stitched fluorescence lifetime image of Convallaria, widefield, 40x objective.
Stitched fluorescence lifetime image of Convallaria, widefield, 40x objective.
Stitched lifetime image of Ranunculus, widefield, 10x objective.
Stitched fluorescence lifetime image of Ranunculus, widefield, 10x objective.

LIFA vTau Short Brochure

LIFA vTau Long Brochure

Publications with and about LIFA FLIM systems

 

Publications with LIFA Toggel (Predecessor of LIFA vTau):

Publications about LIFA Toggel (Predecessor of LIFA vTau):

LIFA Software for Fluorescence Lifetime Imaging Microscopy

LIFA software guides you through your FLIM experiments from start to finish. All Lambert Instruments hardware is integrated seamlessly so you can focus on your experiment. The software takes care of recording the FLIM data and instantly calculates the fluorescence lifetime.

FLIM Settings

Setting up a FLIM acquisition is easy. The Live View images from the camera show you how each of the settings affects the resulting image. Once you’ve configured the FLIM settings you click Record reference/sample to start the image acquisition. You can also record a single image by clicking Snapshot.

LIFA vtau Toggel Camera Settings

 

Light-Source Controls

Switching between wavelengths is done with a single click with the Lambert Instruments Multi-LED and Multi-LASER. Both are integrated seamlessly into the LIFA software.

Light-Source Control

 

Time-Lapse

Follow changes in lifetime with a time-lapse. Simply enter the duration and the interval between lifetime images and see the results immediately in the image viewer and the graphs.

Time-Lapse Settings

 

Data Analysis

Data analysis is easy with automatic lifetime calculation and visualisation. You can overlay the lifetime as a pseudo-color on the image of your sample or use one of the built-in graphs. All graphs are updated continuously during image acquisition and you can choose between displaying one, two or four different graphs.

Lifa vtau toggel Analysis Tab Overview Cropped

 

Lifetime Overlay

Visualize the fluorescence lifetime as a pseudo-color overlay on your sample. You can use the automatic overlay settings or manually adjust the color overlay parameters.

Lifa vtau Lifetime Overlay Option

 

Scatter Plot

Scatter plots show how a value changes over time or as a function of another quantity. This is especially useful for illustrating changes in lifetime in a time-lapse.

Scatter Plot

 

Histogram

Histograms are useful to get a quick overview of the values that are present in your sample and how they are distributed. The histogram chart in LIFA can show lifetime or light intensity.

Histogram

 

Phasor Plot

The phasor plot combines information from both phase and modulation to visualise the contribution of multiple lifetime components in your sample.

Phasor Plot

 

File Format

The Lambert Instruments file format (.fli) is compatible with Bio-Formats. This means you can open .fli files in analysis programs like ImageJ, CellProfiler and Matlab.

 

LIFA API

The LIFA software offers an Application Programming Interface (API) that lets you automate many of its functionalities. This enables endless possibilities for integration with other hardware and software. The LIFA API .Net Assembly can be used in combination with many software platforms, including MatLab, LabVIEW & Python 3 (64-bit).