Leica SP8 Falcon + coherent anti-Stokes Raman Scattering (CARS)
System Specifications:
This is a commercial imaging platform customized to feature the following modalities: confocal fluorescence and two-photon excited fluorescence (TPEF) microscopy, second harmonic generation (SHG), CARS and fluorescence lifetime microscopy. This imaging system was recently acquired through a high-end shared instrumentation grant. Support for this shared resource is provided by the Beckman Laser Institute & Medical Clinic, the Chao Family Comprehensive Cancer Center and the Skin Biology Research Center at UC Irvine. Please, contact Amanda Durkin (afdurkin@hs.uci.edu) and Mihaela Balu (mbalu@hs.uci.edu) if you are interested in using this imaging system.
– InSight X3 laser (dual output: 680-1300 nm, 120 fs; 1045 nm, 170 fs; 80 MHz, Spectra-Physics)
– CW lines and a white-light laser source for confocal fluorescence microscopy – it consists of a high-energy pulsed IR-fiber laser that is fed through a photonic crystal fiber to generate a spectral continuum
– Upright microscopy configuration with a sample staging area that allows for maximum flexibility in terms of sample handling
– Tandem scanner 8kHz
– One internal detector channel (PMT), four internal non-descanned detector channels (GaAsP-HyD) which are specifically designed for maximum throughput and optimized for NLO imaging
– DIVE detection method – allows exquisite control over selecting a particular spectral band while suppressing background contributions
– FALCON detector – one of the fastest FLIM detectors commercially available.
– Frame sizes of up to 8192 x 8192 pixels; features a rapid mosaic stitching algorithm that synchronizes the scanning mirrors with the motorized sample translation stage.
FLAME: Fast, Large Area Multiphoton Exoscope
Home-built multiphoton microscopy (MPM) system for clinical skin imaging
System Specifications:
– Femtosecond fiber laser (100 fs, 780 nm, Carmel780, Calmar)
– Galvo/resonant scanning system
– 2 photomultiplier tube detectors employed for parallel acquisition of SHG and TPEF signals
– Emission filter TPEF channel: 506 – 700 nm, emission filter SHG channel: 390 – 506 nm
– Objective: Olympus 25X, 1.05NA water immersion
– Pixel dwell time max 0.3 μs (effective pixel dwell time: max 2.5 μs)
– Laser power after objective 10-60mW
– Custom software designed for data collection
Commercial multiphoton tomograph for clinical skin imaging (MPTflex, JenLab GmbH, Germany)
System Specifications:
– Laser source: Ti:Sapphire oscillator (MaiTai, Spectra-Physics, Mountain View, CA), tunable 690-1040 nm, sub-100 fs, 80MHz
– Articulated arm with NIR optics and beam scanning module
– Photomultiplier tube detectors employed for parallel acquisition of SHG and TPEF signals
– Emission filter TPEF channel: 410 – 650 nm, emission filter SHG channel: 385 – 405 nm
– Objective: Zeiss 40X, 1.3NA oil immersion
– Pixel dwell time 22 μs
– Laser power after objective 2-50mW
– Software designed for clinical data collection
GSLab: Open-source platform for advanced phasor analysis in fluorescence microscopy
The NLOM has developed an open-source phasor analysis tool available for download:
– Windows installer version.
– GitHub landing page.
– Open-source version for MATLAB.
– Tutorial and sample dataset.
– GitHub for developers.
– UCI landing page.
Zeiss LSM 510 Meta microscopy system
System Specifications:
This imaging system serves as shared resource for the Chao Family Comprehensive Cancer Center and the Skin Biology Research Center at UC Irvine.
– Mode-locked Ti: Sapphire laser (170 fs pulse width, 690-1040 nm tuning range, Chameleon Ultra, Coherent, Inc.)
– Zeiss Axiovert 200M inverted microscope
– 6 lines for confocal scanning 458/477/488/514/532/633 nm
– Up to 4 channels parallel detection
– Scanning speed of up to 5 frames/second for acquisition of 512×512-pixel images
– Motorized X-Y stage with mark and find and tile scan functions and fast piezo objective focus for Z drive with 25 nm smallest
– User-defined ROI.
– Multiple acquisition modes such as spot, line, frame, Z-stack, lambda stack, or time series.
– The polychromatic 32-channel detector (META detector) provides spectral separation of fluorophores within the same sample by spectral fingerprinting and linear unmixing algorithms.
– Multiple acquisition modes such as spot, line, frame, Z-stack, lambda stack, or time series.
– The polychromatic 32-channel detector (META detector) provides spectral separation of fluorophores within the same sample by spectral fingerprinting and linear unmixing algorithms.