Resources

Leica SP8 Falcon + coherent anti-Stokes Raman Scattering (CARS)

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.

System Specifications:
– 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.

Zeiss LSM 510 Meta microscopy system

System Specifications:

– 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.

This imaging system serves as shared resource for the Chao Family Comprehensive Cancer Center and the Skin Biology Research Center at UC Irvine.

Home-built multiphoton microscopy (MPM) system for clinical skin imaging: fast, large area multiphoton exoscope (FLAME)

System specifications:
– fs 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: 450 – 700 nm, emission filter SHG channel: 390 – 410 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