bh FLIM: More than Fluorescence-Lifetime
Imaging
bh FLIM systems record FLIM images of
unprecedented temporal and spatial resolution at an accuracy level close to the
theoretical limit given by photon statistics. But bh FLIM systems do more that
that: The bh FLIM technique is based on a new understanding of FLIM in general.
FLIM is not just considered a way to add additional contrast to microscopy
images. It is considered and designed as a molecular imaging technique. bh FLIM
exploits the fact that the fluorescence decay function of a fluorophore is an
indicator of its molecular environment, and that multi-exponential decay
analysis delivers molecular information, such as the metabolic state of live
cells and tissues, protein conformation and protein interaction, reaction of
cells to drugs and environment parameters, or mechanisms of cancer development
and cancer progression. To reach this target, bh FLIM systems have features not
available in other systems: Compatibility with live-cell imaging, extraordinarily
high time resolution and photon efficiency, capability to split decay functions
into several components, excitation-wavelength multiplexing in combination with
parallel-channel detection, recording of dynamic lifetime effects caused by
fast physiological effects, and simultaneous FLIM/PLIM.
Precision Megapixel FLIM Images
Pixel numbers in
the megapixel range and precision decay data. Enjoy the beauty of the images!

The Ultimate in FLIM Time Resolution and Timing Stability
Electrical time
resolution 3.5ps fwhm. Timing stability better than 0.4ps rms. System IRF
<19 ps fwhm including detector and laser. No need to record an IRF!

High Resolution Decay Data - The Basis of Metabolic FLIM
NADH FLIM:
Metabolic Ratio, lifetimes of unbound and bound NADH

Simultaneous FLIM of NADH and FAD - Tumor Detection by
Metabolic FLIM
Metabolic FLIM, excitation-wavelength multiplexing,
simultaneous imaging of NADH and FAD. Perfect Separation of NADH and FAD.

Protein Interaction - Quantitative FRET Results
Precision FLIM data, double-exponential FRET
analysis: No need to record free-donor lifetime from reference sample.
Left to
right: Classic FRET efficiency, FRET efficiency of interacting donor, amount of
interacting donor, donor-acceptor distance.

Ultra-Fast Decay
Processes
Discover fluorescence-decay processes which
have never been seen before. Below: Mushroom spores, fast decay component of
11 ps.

Below: Melanoma
sample, decay curves of healthy tissue and tumor tissue. The tumor has a fast
decay component of 13 ps.

Autofluorescence
Imaging of Small Organisms
Study environment effects on small
organisms

Triggered Accumulation
of Time Series - Recording of Fast Physiological Effects
Left: Calcium transient in cultured neurons. Right:
Chlorophyll transient.

Online FLIM - Minimum Acquisition Time for Given Photon
Rate
Track tho object of Interest. FLIM with
200 ms Acquisition Time.

Temporal Mosaic FLIM:
Precision Lifetime Analysis of Moving Objects
Metabolic FLIM on the moving leg of a water
flee. bh Temporal Mosaic FLIM with subsequent Image Segmentation. Precision
decay curve shown lower right.

Simultaneous FLIM / PLIM
Record the metabolic state of cells in
dependence of oxygen concentration.

Multi-Wavelength Detection
Explore the unexplored: Simultaneous Detection
in 16 Wavelength Channels

For more information please see:
1.
W. Becker, The bh TCSPC handbook, 9th edition.
Becker & Hickl GmbH (2021), available online on www.becker-hickl.com. Please contact bh for printed copies.
2. Becker & Hickl GmbH, FLIM Systems for Laser Scanning
Microscopes. Overview brochure, available on www.becker-hickl.com
3. W. Becker (ed.), Advanced time-correlated single photon counting
applications. Springer, Berlin, Heidelberg, New York (2015)
4. Becker Wolfgang, Suarez-Ibarrola Rodrigo, Miernik Arkadiusz, Braun
Lukas, Metabolic Imaging by Simultaneous FLIM of NAD(P)H and FAD. Current
Directions in Biomedical Engineering 5(1), 1-3 (2019)
5. W. Becker, Bigger and Better Photons: The Road to Great FLIM
Results. Education brochure, available on www.becker-hickl.com.
6.
Becker & Hickl GmbH, SPCImage next
generation FLIM data analysis software. Overview
brochure, available on www.becker-hickl.com
7. Becker & Hickl GmbH, Simultaneous Phosphorescence and
Fluorescence Lifetime Imaging by Multi-Dimensional TCSPC and Multi-Pulse
Excitation. Application note, available on www.becker-hickl.com
8. S. Kalinina, V. Shcheslavskiy, W. Becker, J. Breymayer, P. Schäfer,
A. Rück, Correlative NAD(P)H-FLIM and oxygen sensing-PLIM for metabolic
mapping. J. Biophotonics 9(8):800-811 (2016)
Becker & Hickl GmbH
Nunsdorfer Ring 7-9
12277 Berlin, Germany
Tel. +49 30 212 800 20, Fax. +49 30 212 800 213
email: info@becker-hickl.com,
www.becker-hickl.com
