Imaging Core Equipment

Yokogawa CSU-W
E015B -- The Yokogawa CSU-W1 spinning disk confocal system is coupled to a Nikon Eclipse NI-E upright microscope with two Andor EM CCD detectors for simultaneous two-channel imaging. Three laser lines are available, 488 nm 561 nm and 640 nm. The Yokogawa CSU-W1 has two water-immersion objectives: a Nikon Apo LWD 25X/1.1NA, and a Nikon Fluor 60X/1.0NA, both with 2mm working distance. This instrument is designed for fast image collection, with rates in excess of 50 fps. With its widened-diameter Nipkow disk, the CSU-W1 provides at least four times greater field of view than comparable systems, increasing the likelihood of capturing Ca2+ transients and other events in precious tissue, and increasing throughput. The wider disk also allows wider pinhole spacing, which decreases flare or crosstalk between pinholes, significantly improving image quality. Two detectors can be used simultaneously with emission filters for 505-545nm and 580-650nm, or 505-545nm and 665-705nm.
Zeiss LSM-7 Dedicated Multiphoton
Given E015C -- The Zeiss LSM-7 Dedicated Multiphoton has four non-descanned PMT detectors, two of which make up the highly sensitive Zeiss gallium arsenide phosphide BiG detector. Emission filters include 485nm and below, 500-550nm, 570-645nm, and 640-710nm, as well as a DIC (differential interference contrast) transmitted detector. Multiphoton excitation is generated by a Coherent Chameleon Vision II Titanium Sapphire pulsed IR laser, with dispersion compensation, and tunable from 720-950 nm. The stage is a motorized Prior Z-deck, which is fully integrated into the Zeiss Zen software. A 20X Plan Apo/1.0 NA DIC VIS-IR water immersion lens is the main objective. The system frame is a Zeiss Axio Examiner, specifically optimized for multiphoton microscopy.
Nikon C2 Confocal Microscopy System
Given E015D -- The Nikon C2 Confocal Microscopy System is a general use point-scanning confocal. The C2 system is based on a Nikon Ti-E inverted platform equipped with Plan Apo and Plan objective lenses with DIC capability, including a 10X/0.45 NA, 20X/0.75NA, 40X/1.0 NA oil and 60x/1.4 NA oil. Solid-state, fiber coupled lasers provide excitation at 405nm, 488nm, 561nm and 640nm. Three photomultiplier tubes can capture fluorescent images simultaneously, along with one dedicated transmitted detector. Emission filters are set at 427-463nm, 500-550nm, 575-625nm and >660nm. The 405nm and 640nm channels must be collected separately.
Delta Vision RT Restoration Microscope
Given E015E -- DeltaVision RT Restoration Microscope: The Applied Precision DeltaVision RT is coupled to a DIC-equipped Olympus IX70 and a Photometrics CoolSnap CCD. Objectives include 20X/.5NA dry, 20X/.8NA oil, 40X/1.35NA oil, 60X/1.4NA oil and 100X/1.4NA oil. This system has a temperature and CO2-controlled incubator, and a high-precision, computer-controlled motorized stage for tiled and multi-point data collection. Fast-sequential emissions can be collected at 432-482nm, 507-543nm, 580-631nm, and 669-741nm. Fast deconvolution of image data is accomplished with a dedicated server, separate from the acquisition computer.
PTI Monochromator
Given C456 -- The PTI Monochromator calcium imaging system is based on a Nikon Eclipse E600FN upright microscope platform coupled with a PTI Monochromator and Photometrics Evolve EMCCD. We have two physiology dipping lenses, a Nikon 60X/1.00N DIC and a Nikon 40X/0.80 DIC. The monochromator allows for precise excitation wavelengths and fast wavelength switching, accommodating a wide range of Calcium signaling and other ratiometric modalities. Currently it is used for both Fura-2 and Fluo-4 labeled live specimens.
TIRF

Given B312 -- The Nikon TIRF microscope is based on a Nikon TE2000-E Microscope. It is a “through the lens” TIRF, using a Nikon Plan Apo TIRF 60X/1.45 Oil ∞/0.10-0.22 lens coupled to a Coherent Sapphire 488 nm 100 mW laser. The camera is a Photometrics Cascade 512B with on-chip multiplication gain for low- emitting samples. For longer term imaging studies, the system is equipped with a Nikon T-PFS Perfect Focus Unit to maintain the correct imaging plane. All the systems are integrated through Nikon Elements software, with provision to trigger electrophysiology equipment via TTL as well. This is currently in Pharmacology space and will be moved into Core space after the renovation.

Noran OZ

Given E025A -- Noran OZ The Noran single-beam scanning confocal imaging system uses a 50mW Coherent Sapphire 488 nm solid state CW laser coupled to a Nikon Eclipse TE 2000-U inverted microscope equipped with 20x and 60x water immersion objectives and DIC optics.The system was retrofit by Prairie Technologies with a modern scan control system and PC interface, and recently upgraded further with a Windows 7 operating system. This is a very fast single-beam confocal laser scanning microscope. The high speed is achieved by using an acousto-optic deflector. It is capable of a 2-D scan rate up to 480 images per second and is ideal for most high-speed ion imaging applications. This system is best suited to measure fast transient calcium events (Ca2+ sparks). In addition, it has a line scan capability with a maximum of 60 ps/line, which allows for the resolution of very fast (ms) events. We have modified this instrumentation to enable simultaneous acquisition of rapid calcium events and membrane potential from endothelium or smooth muscle of pressurized arterioles. This required the addition of a motorized micromanipulator (Sutter MP 285) and a second computer for acquisition, including the software (PClamp; Axon Instruments) and hardware (Digidata 1322A, Axopatch 200A and Axoclamp 2B, Axon Instruments). This enables us to detect rapid calcium events and electrical events that are coupled.