Three-Channel Laser Scanning Confocal Fluorescence, Photoactivation/Uncaging, Brightfield

This three-channel laser scanning confocal is designed on a Nikon Eclipse Ti inverted microscope as a modular system based on a core galvanometer imaging module. Researchers can customize their experiments with an automated 9-pinhole aperture (pinhole size ranging from 30 mm – 2000 mm), filter wheels for each of the three detection channels, and 4-position automated dichroic holders. This system has a photoactivation module along with uncaging/photoactivation optics for simultaneous imaging and photoactivation/uncaging experiments. The motorized stage allows for the automated collection of data at multiple sample locations.

Multiphoton Laser Scanning, Large Field-of-View Forward SHG and Brightfield

CAMM is an inverted multiphoton laser scanning system best suited for histological samples for coregistration of large field-of-view forward detected second harmonic generation photons and brightfield built on an ASI RAMM frame that uses a tunable Coherent MIRA900 laser excitation source (S/N 96994) for imaging from 700-1000 nm pumped by a Coherent VerdiV10 (S/N V10-G9920) with XY scanning control by 

Multiscale Quantitative Ultrasound (QUS), Forward and Backward SHG, Enhanced Backscatter, Optical Coherence Tomography (OCT), Brightfield

LINK is an instrumentation collaboration between the Medical Engineering group in the Morgridge Institute for Research and LOCI investigators that combines quantitative ultrasound (QUS), forwards and backwards scattered second harmonic generation microscopy (SHG), and two optical scattering techniques, enhanced backscatter and optical coherence tomography (EBS and OCT, respectively) to analyze collagen organization in intact animal and human tissues.

Brightfield

The dual head teaching microscope is an upright microscope located in the main LOCI office for histological purposes. The image software associated with this instrument is Olympus CellSens Standard 1.16.

 

 

Laser Scanning Confocal Fluorescence, Fluorescence Recovery After Photobleaching (FRAP)

The Opterra Multipoint Scanning Confocal Microscope is an inverted laser scanning confocal microscope being developed in collaboration with LOCI for improvements in speed, software analysis, photomanipulation and fast spectral collection. The current prototype at LOCI can collect 8 spectral channels at 4 frames a second using an Amici prism incorporated into a multi-pinhole confocal to allow dynamic spectral scanning for live cell imaging in real-time.  The software-controlled aperture design uses a combination of one-dimensional pinhole arrays for maximum resolution with half of the crosstalk, as well as aperture slits for high-speed acquisition. This combination results in sharper images, higher resolution, greater depths, and faster speeds than spinning disk confocal.

Fluorescence Lifetime Imaging (FLIM), Multiphoton Laser Scanning Fluorescence, Brightfield, Optical Ablation

The Optical Workstation (OWS) is an inverted multiphoton laser scanning system that uses a Nikon TE300 inverted microscope frame, ASI MS-2000 XYZ stage and WiscScan interface for time-domain fluorescence lifetime imaging (FLIM), live tissue imaging, fluorescence, and second harmonic generation.  

Multiphoton Laser Scanning, Backscattered SHG, Epifluorescence

OWS II is an inverted multiphoton laser scanning instrumentation development platform with backscattered second harmonic generation (SHG) and epifluorescence capabilities that uses a Nikon Eclipse Ti-U inverted base equipped with an epifluorescent excitation lamp, Spectra-Physics Tsunami Model 3941-X1BB (S/N 1561G) Tsunami Series Ti-Saph laser and a Millennia Pro s-Series Spectra Physics (S/N E2362)

Fluorescence Lifetime Imaging (FLIM), Multiphoton Laser Scanning Fluorescence, Brightfield

Spectral lifetime imaging microscopy (SLIM) simultaneously measures fluorescence lifetime and spectra for intrinsic and extrinsic fluorophores. SLIM is fitted on a Nikon Eclipse TE2000U base.  An excitation laser is tunable from approximately 700-980 nm.   The Hamamatsu H7422 gathers traditional multiphoton intensity images and fluorescent lifetime data.  The Hamamatsu MCP is an extremely sensitive detector used for fluorescent lifetime imaging.  The system also contains a galvanometer scanning system from Cambridge Technologies.

Three-Channel Multiphoton Laser Scanning, Fluorescence Lifetime Imaging (FLIM), Optional Substage Forward and Backward SHG, Brightfield, Intravital

The Bruker Ultima IntraVital is a four-channel multiphoton laser scanning microscope with second harmonic generation (SHG) and fluorescence lifetime capabilities designed for imaging living things in depth, with up to thirteen inches of space beneath the objective to allow live imaging of medium-sized organisms or tissue.  A brightfield light can be used to locate an area of interest, and a substage detector allows for direction-dependent polarization experiments.

Inverted Tissue Culture Fluorescence, Brightfield

Zeiss Axiovert 25 CFL inverted light microscope for routine analyses of cell and tissue cultures in transmitted and fluorescent reflected light.

 

 The fluorescence setup has the following filter sets:

 

FL Filter Set 02, --example fluorophore DAPI

Zeiss filter set 02 excites at 365 nm. and allows emission at 420 nm.

FL Filter Set 09, --example fluorophore GFP

Zeiss filter set 09 excites between 450-490 nm. and allows emission at 515 nm.

FL Filter Set 15, --example fluorophore Rhodamine