|Instrument||Model #||Description||Excitation Sources||Detectors||Signup|
|3D Niche @ WID||WID Town Center 3D Niche||
The 3D Niche houses a microscope, a macroscope, and a 3D printer. The high-resolution Nikon microscope, an ECLIPSE 90i, is capable of DIC and fluorescence and uses the objectives listed below. Data are acquired using NIS-Elements software.
The lower-resolution Nikon macroscope is made up of the Mi Macro Imaging Station and a moveable stage built by Prairie Technologies. This system provides detailed pictures of larger samples and can be used in conjunction with the microscope to more fully study an organism or sample.
The 3D printer is used to build 3D models of any image provided via software. These images can be graphic designs from artists or pictures of objects taken from the microscope itself.
|4D Inverted Fluorescence Microscope|
|Boom Dissecting Stereo Microscope|
|Dual Head Teaching Histology Microscope||BX60||
The Dual Head Teaching Scope is an upright microscope located in the main LOCI office, and it is especially useful for histology.
The objectives available on the microscope are:
|GFP Dissecting Stereo Microscope|
|LOCI Confocal @ BOCK||Multi-purpose Three Channel Confocal||
This Multi-purpose Three Channel Confocal was designed and built by Prairie Technologies on a Nikon Eclipse Ti inverted microscope. The confocal microscope is a modular system based on the core galvanometer imaging module found in other Prairie confocals.
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. This confocal is also capable of reflectance confocal imaging with the 515 nm laser.
|LOCI Confocal Development System||SP2 AOBS||
Leica SP2 AOBS system mounted on DMIRE2 inverted microscope. Multiple laser lines allows for excitation of a wide range of fluorphores, and simultaneous multi-spectral scanning is possible. The microscope also employs a galvanometer z-stage, transmitted light detector and manual condenser with DIC optics.
The OPO (Optical Parametric Oscillator) System is a multiphoton long-wavelength scanning microscope system under development by LOCI. It is a hybrid design of fabrication systems including the Fluoview imaging system and uses Wiscan imaging software. Forward detection can be used to measure second harmonic generation (SHG) and third harmonic generation (THG). The system will also have adaptive optics and lifetime measurement capabilities. The OPO System will have a wide range of uses including broadband excitation, label-free imaging, polarization control, and deep imaging.
|Opterra Multipoint Scanning Confocal Microscope||
Spectral Swept-Field Confocal
The Opterra Multipoint Scanning Confocal Microscope is 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. The short acquisition times minimize photobleaching and phototoxicity, and this cell-friendly 4D instrument is ideal for studies including protein localization and trafficking, microtubule dynamics, mitosis, and FRAP.
[caption] Mitosis in Xenopus laevis embryo
The cell membrane is labeled with targeted mTagBFP, microtubules are labeled with eGFP-tubulin, and chromatin is visualized with mCherry-Histone H2B. The image was captured with 500 ms exposure time. A video of the mitosis is provided in supplementary movie 2. The image was captured with filter configuration 2 (see section "System Dichroics and Emission Filters") with a 60x oil objective and excitation at 405 nm, 488 nm, and 561 nm.
Velten A., Fong J. J., White J. G., Vogt W., Plavicki J., Larson M. E., Squirrell J. M., Harvie E.A., Huttenlocher A., Bement W. M., Mackie T. R., Eliceiri K. W. (2014). Hyperspectral multipoint high-speed confocal microscopy. Manuscript submitted for publication.
|Prairie Swept-Field Calendar|
|Optical Work Station Multiphoton||OWS||
This is a custom built multiphoton laser scanning system built around a Nikon TE300 inverted microscope. The system features dual galvos for simultaneous imaging and photomanipulation, three lasers and time-domain fluorescence lifetime.
The OWS is the system most often used by collaborators. Experiments often run on the OWS include collagen imaging of tumor sections using second harmonic generation, lifetime imaging, and imaging of autofluorescent molecules such as NADH.
Under a UW-Milwaukee and UW-Madison Biophotonics partnership the Raicu group is developing the OptiMiS multiphoton microscope. This is a novel multiphoton microscopy with spectral resolution for with spectral resolution forr spectrally resolving heterogeneous signals in live cell imaging. A specific focus is fluorescence resonance energy transfer (FRET) studies.
Using the TPM-SR, as well as a novel theory of FRET, the Raicu group has determined the stoichiometry, geometry and spatial distribution of the oligomeric complexes of a yeast G protein-coupled receptor, Ste2p.
|OptiMiS (Ultima) Calendar|
|Prairie Ultima Intravital Multiphoton||Prairie Ultima IV||
The IV Multiphoton was built by Prairie Technologies in Middleton, WI. It is especially useful for in vivo research but can be used for a variety of multiphoton applications. The stage is separate from the microscope setup and can be removed to allow for additional clearance under the lenses; up to 13 inches of space under the objective lens allows for visualization of medium-sized organisms. A brightfield light can be used to locate an area of interest, and a substage detector allows for direction-dependent polarization experiments.
|Prairie Intravital Calendar|
|Spectral Lifetime Multiphoton Microscope||Spectral Lifetime Imaging Microscope (SLIM)||
Spectral lifetime imaging microscopy (SLIM) simultaneously measures fluorescence lifetime and spectra for intrinsic and extrinsic fluorophores. This technique can discriminate between multiple fluorophores and provide information about the microenvironment.
The SLIM system at LOCI is fitted on a Nikon Eclipse TE2000U base. An excitation laser is tunable from approximately 700-980 nm. Three detectors allow for recording of emitted fluorescence.
Ultra is a custom built non-linear microscopy imaging system using the Olympus BX61 as its base. The system has 3 independent channels allowing simultaneous collection of forward and backward Second harmonic generation (SHG) signals as well as third harmonic generation (THG). The laser source is a tunable Chameleon/OPO system allowing imaging from 780-1230nm.
|Zeiss Inverted Tissue Culture Fluorescence Microscope||Axiovert 25||
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
Zeiss filter set 15 excites at 546/12 nm. and allows emission at 590 nm.
The scope is also equipped with a Object Marker w/Felt Tip W0.8