Laboratory for Fluorescence Dynamics

A NIH research center for biomedical fluorescence spectroscopy at the University of California, Irvine

Spotlights

Selected activities of the Laboratory for Fluorescence Dynamics (LFD) in random order.

Globals Software

Screenshot of Globals for Images

Globals software, originally developed in the 1980s at the LFD for the analysis of multiple files from spectroscopy, is now available for the Windows operating system (Globals for Spectroscopy) and for image analysis using the RICS, N&B, and Phasor approaches (Globals for Images aka SimFCS).

The programs are available for download.

Weber Symposium

Photo of Weber Symposium

The 10th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine was held in Buzios, Brazil, on May 28 - June 2, 2017.

The symposium honors the fundamental and far-reaching contributions of Gregorio Weber. It provides an overview of modern fluorescence methodologies and applications in the biological and medical sciences.

Recordings of many lectures of previous symposia are available.

Phasor Plot

Screenshot of Phasor Plot Analysis

The phasor plot is a "fit free", interactive approach for analyzing fluorescence lifetime-resolved images acquired in the time- or frequency-domain.

It is used to determine Förster Resonance Energy Transfer (FRET) distributions and for mapping of multiple components in cells and tissues.

The phasor plot analysis is part of the Globals for Images software.

Frontiers in Biological Fluorescence 2006

Photo of Enrico Gratton

The symposium to honor Enrico Gratton on the occasion of his 60th birthday and 30-years of service to the University of Illinois was held on May 20, 2006 at the Loomis Laboratory of Physics, University of Illinois at Urbana-Champaign.

Presentation slides and audio recordings are available.

Services and Resources

Photo of the laboratory

The LFD provides a state-of-the-art laboratory for fluorescence measurements with technical assistance to visiting scientists:

  • Spectroscopy, steady-state and time-resolved
  • Microscopy, capable of FCS, FLIM, RICS, GP and particle tracking
  • Biochemistry and cell culture
  • Data analysis and visualization

People

Photo of LFD Members

About two dozen people are currently working at the LFD.

More than thirty students have graduated out of the LFD since its foundation in 1986.

Many former members are distinguished scientists in academia and industry.

The resource advisory committee meets annually to assist the LFD.

University of California, Irvine

The Natural Sciences II building at UCIrvine

In 2006 the LFD relocated from the University of Illinois at Urbana-Champaign to the University of California, Irvine, Biomedical Engineering Department, where it occupies a ~8000 sq. ft. facility in the interdisciplinary Natural Sciences II building.

Take a picture tour through the new facility.

Tribute to Gregorio Weber

Photo of Gregorio Weber

In tribute to the outstanding contributions of Gregorio Weber (1916-1997) to the field of fluorescence, the LFD organizes and sponsors the:

Particle Tracking

Rendering of a microvilli

The circular scanning particle tracking technique developed at the LFD allows tracking the 3D position of fluorescently labeled molecules, organelles, or viruses in live cells.

It determines and follows the center of mass of a fluorescence signal at nanometer and millisecond resolution using a laser scanning confocal microscope.

The technique has recently been extended to allow dynamic 3D imaging of fluorescently labeled surfaces in live cells and tissues.

Publications

Picture of Books and Journals

The LFD database contains more than 2100 articles, abstracts, patents and theses by members of the LFD, and also selected publications of interest to the fluorescence community.

SPIM

SPIM 3D reconstruction

Single plane illumination microscopy (SPIM) in an upright configuration allows for fast 3D imaging of single cells grown on glass coverslips, and of large, multicellular organisms such as zebrafish embryos.

The custom-built SPIM system at the LFD can simultaneously image two color channels, e.g. the nucleus (blue, H2B-EGFP) and mitochondria (green, TMRE) of live CHO-K1 cells (shown).

Weber Prize

Photo of Weber Prize ceremony

The Gregorio Weber International Prize in Biological Fluorescence is awarded by the LFD for research related to a doctoral or equivalent dissertation. All fields of biological fluorescence (experimental, theoretical, or applied) are eligible. The award is conferred approximately every three years.

The 2017 Weber Prize winner received a cash award of $2500 and an invitation to the 10th International Weber Symposium.

Biophysical Society Meeting

Biophysical Meeting booth poster

The 62nd Annual Meeting of the Biophysical Society will be held on February 17-21, 2018, in San Francisco, California.

Many lab members attend the meeting and present posters and talks.

The Biological Fluorescence Subgroup is chaired by Enrico Gratton.

The LFD organizes an exhibitor booth, where visitors are informed about the LFDs research and service facilities. The LFD Workshop and Weber Symposium are advertised and Globals software is demonstrated.

Undergraduate Student Initiative

USIBR logo

The Undergraduate Student Initiative for Biomedical Research (USIBR) exposes underrepresented community college students to cutting-edge technology used to study living organisms. The program provides lectures and hands-on training on the basics of microscopy, optics, imaging methods, and computational data analysis.

Research and Development

Advances in Logo Design

The LFD designs, tests, and implements advances in the fluorescence technology of hardware, software, and biomedical applications. Core projects:

  • Hardware and software for the microscope environment
  • Spatio-temporal fluctuation correlation spectroscopy
  • 3D particle tracking and distance measurements
  • Fluorescence lifetime-resolved imaging
  • Globals data analysis software

DIVER

DIVER

Deep Imaging Via Enhanced-photon Recovery (DIVER) is a microscope system developed at the LFD capable of fluorescence imaging of tissue with up to 4 mm in depth with micron resolution.

A unique, wide surface detector allows for an 8 fold increase in the depth compared to conventional two-photon imaging. The system has been used to image intestine, heart, kidney, liver and lung tissue. It is also very efficient in Second Harmonic Generation (SHG) imaging and can be combined with Fluorescence Lifetime Imaging (FLIM).

See US Patent 8692998.

FlimBox

FlimBox hardware

The FlimBox device developed at the LFD enables ultra high bandwidth, multi-channel data collection for fluorescence lifetime-resolved imaging (FLIM) from a pulsed laser scanning confocal microscope.

The device is implemented using a commercial, low cost field programmable gate array (FPGA) chip and uses the digital frequency domain (DFD) approach.

Collected data can be analyzed directly using the multi-harmonic phasor plot.

Giant Unilamellar Vesicles

A Giant Unilamellar Vesicle

Fluorescently labeled Giant Unilamellar Vesicles (GUVs) are used as a model system for studying fluctuations in membranes, lipid-lipid interactions, and protein-membrane interactions.

Confocal fluorescence images of GUVs (left) are analyzed for fluctuations and polarization (right) using Globals for Images software.

Introductory Course

Rendering of a cell

The Introductory Course in Fluorescence Techniques, Spectroscopy and Microscopy was held on March 23 - 25, 2009. On the agenda:

  • Fluorescence spectroscopy basics: probes, instrumentation, spectra, quantum yield, anisotropy, energy transfer, lifetime-resolved, FCS
  • Fluorescence imaging: confocal and 2-photon, correlation methods, number and brightness analysis
  • Theoretical lectures and optional hands on laboratory training

Tissue Imaging

Bone tissue

The DIVER microscope system developed at the LFD can image large tissue areas in various imaging modes.

For example, a femur slice from a SMRTmRID mouse is shown in bright field (top), second harmonic generation (SHG, middle), and fluorescence lifetime imaging (FLIM, bottom) modes. The FLIM image is phasor mapped to segment Collagen I (red) from Collagen III (turquoise) rich regions.

See Sci Rep. 2015; 5: 13378.

Lecture Recordings

Screenshot of Flash video

Many lectures organized by the LFD are audio/video recorded and available for on-demand viewing:

LFD Workshop

Photo of LFD Workshop

The 12th LFD Workshop in Advanced Fluorescence Imaging and Dynamics was held on October 23 - 27, 2017. On the agenda:

  • FCS, FLIM, FRET, RICS, N&B, pCF, PCH, DIVER, SPIM, iMSD, ipCF, phasors, and particle tracking
  • Theoretical lectures
  • Hands on laboratory training
  • Data analysis and simulations using Globals software

Lecture notes and recordings are available.

Young Fluorescence Investigator

Photo of Dr. Digman

February 2012: Dr. Michelle Digman wins the Young Fluorescence Investigator Award, sponsored by HORIBA Jobin Yvon Inc.

Dr. Digman was recognized at the Biophysical Society Annual Meeting 2012 for significant advancements and contributions in or using fluorescence methodologies. More...