Information for grant applications

Light Microscopy Core Facility
The Light Microscopy Core Facility (LMCF) offers a wide range of confocal and conventional fluorescence microscopes and image analysis resources. As a centrally funded shared resource, the core’s aim is to offer affordable and efficient access to standard and advanced imaging instrumentation for users of all levels of experience and from any discipline across the Duke University and the Duke Medical Center campuses.

Facilities
The LMCF is housed in over 2600 square feet of space located in four clusters across the research campus to facilitate easy access to the microscopy resources. Each of the four clusters has at least one confocal microscope, one conventional fluorescence microscope, one microscope with incubation capabilities, one incubator for temporary sample storage, and one image analysis workstation. All locations and resources are open to everybody at Duke.

Equipment
The core facility has twenty-one microscopes of various configurations and modalities, as well as six image analysis workstations and other resources, allowing users to image and analyze a large range of samples at various conditions. Included in the microscopy systems are one multiphoton system with fluorescence lifetime measurement capabilities, nine confocal microscopes, one total internal reflection fluorescence (TIRF) microscope, seven widefield fluorescence microscopes, one laser capture microdissection microscope system, and two stereoscopes and dissecting scopes. Of the microscope systems listed, nine of the systems have environmental controls for the incubation of samples during imaging. In addition to the microscope systems, the core facility maintains six workstations for image processing and analysis of collected images by registered core users.

Multiphoton Microscopes
Olympus FV1000 Multiphoton System: An Olympus BX61WI upright microscope with a tunable Chameleon Ti:Saphire femtosecond laser, four high efficiency non-descanned photomultiplier tube (PMT) detectors, two objectives (25x/1.05NA water, 25x/0.9NA ScaleView immersion), and a PicoHarp 300 counting electronics system for fluorescence lifetime imaging microscopy (FLIM).

Confocal Microscopes
Andor XD Revolution Inverted Spinning Disk Confocal Microscope: An Olympus IX81 inverted microscope with an Andor AOTF controlling 405, 445, 488, 515, and 561 nm laser lines, a Yokogawa CsuX-1 5000 rpm spinning disk, an Andor Ixon3 897 EMCCD camera and an OrcaER camera, and six installed objectives (10x air, 20x air, 40x air, 60x silicon, 60x water, 100x oil).
Leica SP5 Inverted Confocal Microscope: A Leica DMI6000CS inverted microscope with 405, 458, 477, 488, 514, 561, 594, and 633 nm laser lines, an AOBS based system, conventional and resonant scanners, three cooled photomultiplier tube (PMT) spectral detectors, five installed objectives (10x/0.40NA air, 20x/0.70NA air, 40x/1.25NA oil, 63x/1.2 water, 100x/1.4NA oil), and an environmental chamber for live cells.
Leica SP5 Upright Confocal Microscope: A Leica DMI6000CS upright microscope with 405, 458, 476, 488, 514, 561, and 633 nm laser lines, conventional and resonant scanners, four cooled photomultiplier tube (PMT) spectral detectors, and five installed objectives (10x/0.40NA air, 20x/0.70NA multi-immersion, 40x/1.25NA oil, 63x/1.2 water, 100x/1.4NA oil).
Leica SP8 Upright Confocal Microscope: A Leica DMI6000CS upright microscope with 405, 458, 488, 514, 561, and 633 nm laser lines, conventional and resonant scanners, two photomultiplier tube (PMT) and two high sensitivity hybrid (HyD) spectral detectors, and seven installed objectives (2.5x/0.07NA air, 5x/0.15NA air, 10x/0.40NA air, 20x/0.70NA multi-immersion, 25x/0.95NA water, 40x/1.3NA oil, 100x/1.4NA oil).
Zeiss 510 LSM Inverted Confocal Microscope: A Zeiss Axio Observer inverted microscope with 405, 458, 477, 488, 514, 561, 594, and 633 nm laser lines, three photomultiplier tube (PMT) detectors, and six installed objectives (10x/0.30NA air, 20x/0.80NA air, 40x/0.95NA air, 40x/1.3NA oil, 63x/1.4 oil, 100x/1.4NA oil).
Zeiss 510 LSM Upright Confocal Microscope: A Zeiss Axioplan2 upright microscope with 351, 364, 458, 477, 488, 514, 543, 594, and 633 nm laser lines, three photomultiplier tube (PMT) detectors, and five installed objectives (10x/0.30NA air, 25x/0.80NA multi-immersion, 40x/1.3NA oil, 63x/1.2NA water, 63x/1.0 dipping, 100x/1.4NA oil).
Zeiss 710 LSM Inverted Confocal Microscope: A Zeiss Axio Observer inverted microscope with 405, 458, 477, 488, 514, 561, 594, and 633 nm laser lines, two photomultiplier tube (PMT) spectral detectors and a 32-element spectral array detector, five installed objectives (10x/0.45NA air, 20x/0.80NA air, 40x/1.3NA oil, 63x/1.4 oil, 100x/1.4NA oil) , and an environmental chamber for live cells.
Zeiss 780 LSM Inverted Confocal Microscope: A Zeiss Axio Observer Z1 inverted microscope with 405, 458, 477, 488, 514, 561, and 633 nm laser lines, two photomultiplier tube (PMT) spectral detectors (one cooled) and one high quantum efficiency (QE) 32-element spectral array GaAsP detector, and five installed objectives (10x/0.3NA air, 20x/0.80NA air, 40x/1.4NA oil, 63x/1.4 oil, 100x/1.45NA oil).
Zeiss 780 LSM Upright Confocal Microscope: A Zeiss Axio Examiner upright microscope with 405, 458, 477, 488, 514, 561, and 633 nm laser lines, two photomultiplier tube (PMT) spectral detectors (one cooled) and one high quantum efficiency (QE) 32-element spectral array GaAsP detector, and four installed objectives (10x/0.45NA air, 20x/0.80NA air, 20x/1.0NA water, 63x/1.4 oil).

Total Internal Reflection Fluorescence (TIRF) Microscopes
Leica AM TIRF MC Microscope System: A Leica DMI6000 B inverted microscope with a Leica AM TIRF module and a Leica AOTF controlling 405, 488, 561, and 635 nm laser lines, a Hamamatsu imagEM EM-CCD camera, four installed objectives (10x/0.4NA air, 20x/0.7NA air, 40x/0.75NA air, 100x/1.46NA oil), and an environmental chamber for live cells.

Widefield Fluorescence Microscopes
DeltaVision Elite Deconvolution Microscope: An Olympus IX-71 inverted microscope with an InsightSSI 7 channel solid state illumination source switching DAPI, CFP, GFP/FITC, YFP, TRITC, mCherry/AF594, and Cy5 filter cubes, a Coolsnap HQ2 high resolution CCD camera and an Evolve EM-CCD camera, six installed objective (10x/0.4NA air, 20x/0.7NA air, 40x/0.65-1.35NA oil, 60x/1.42NA oil, 60x/1.2NA water, 100x/1.4NA oil), and an environmental chamber for live cells.
Leica DMI 4000 Inverted Widefield Fluorescence Microscope: a Leica DMI 4000B inverted microscope with an X-CITE Series 120Q light source and GFP, YFP, Rhodamine, and Cy5 filter cubes, a Photometrics CoolSNAP HQ2 cooled camera, four installed objectives (5x/0.12NA air, 20x/0.4NA air, 40x/0.6NA air, 100x/1.3NA oil), and a microinjection system consisting of two Eppendorf TransferMan NK2 micromanipulators and an Eppendorf FemptoJet microinjection system.
Olympus VivaView FL Incubator Microscope: an Olympus VivaView FL inverted microscope with an X-CITE eXacte light source and CFP, GFP, YFP, and Red filter cubes, a cooled CCD camera, two objectives available (20x/0.75NA air, 40x/0.95 air) with 0.5x and 2.0x auxiliary magnification available, and an incubator sample chamber for live cells.
Zeiss Axio Imager Widefield Fluorescence Microscope: a Zeiss Axio Imager.Z2 upright microscope with a Zeiss HXP 120 metal halide fluorescence illumination and a LED transmitted light illunination, filter cubes for bright field, DIC, DAPI, CFP. GFP, YFP, Rhodamine, Texas Red, and Cy5, high resolution Axiocam 506 color and Axiocam 506 mono cameras, six installed objectives (5x/0.16NA air, 10x/0.45NA air, 20x/0.80NA DIC air, 40x/0.90NA DIC air, 63x/1.4NA DIC oil, 100x/1.4NA DIC oil), Apotome.2 for optical sectioning, and motorized stage for tile imaging and stitching.
Zeiss Axio Observer Widefield Fluorescence Microscope: A Zeiss Axio Observer A1 inverted microscope with a Zeiss HBO arc lamp and DAPI, CFP, Green, YFP, Red, Cy5, and Cy7 filter cubes, a Hamamatsu Orca ER camera, and six installed objectives (5x/0.16NA air, 10x/0.3 air, 20x/0.5NA air, 40x/0.75NA air, 63x/1.4NA oil, 100x/1.4NA oil).
Zeiss Axio Observer (Live Cell Station): A Zeiss Axio Observer Z1 inverted microscope with an X-CITE 120XL light source and DAPI, CFP, GFP, YFP, and Red filter cubes, a Hamamatsu Orca ER high resolution CCD camera and a QuantEM EM-CCD camera, six installed objectives (5x/0.16NA air, 10x/0.3 air, 20x/0.8NA air, 40x/0.75NA air, 63x/1.4NA oil, 100x/1.46NA oil), and an environmental chamber for live cells.
Zeiss Axiovert 200 M (Live Cell Station 2): A Zeiss Axiovert 200 motorized inverted microscope with an X-CITE 120XL light source and CFP, GFP, YFP, and Texas Red filter cubes, an Orca ER CCD camera, six installed objectives (2.5x/0.075NA, 5x/0.15NA air, 10x/0.5 air, 20x/0.4NA LD air, 40x/0.85NA air, 63x/1.4NA oil), and an environmental chamber for live cells.

Laser Capture Microdissection (LCM) Microscopes
Zeiss Laser Microdissection and Capture Microscope: a Zeiss Axio Observer Z1 inverted microscope with an X-CITE fluorescence light source and DAPI, Green Texas Red filter cubes, an AxioCam MRm monochrome camera and an AxioCam ICc1 color camera, five installed objectives (5x/0.12NA air, 10x/0.25NA air, 20x/0.4NA LD air, 40x/0.60NA LD air, 63x/0.75NA LD air), and a PALM MicroBeam 4.2 system with a Robomover Z device for excising and harvesting samples in a non-contact manner.

Stereoscopes and Dissecting Scopes
Olympus SZ61 Dissecting Scope: an Olympus SZ61 dissecting scope with a zoom range from 0.67x to 4.5x, a field of view up to 65 mm, and a working distance of 110 mm.
Zeiss Lumar.V12 Stereoscope: a Zeiss Lumar.V12 stereoscope body with a Zeiss HBO100 fluorescence arc lamp and Green and Red filter cubes, an AxioCam MRM monochrome camera and an AxioCam ICc3 color camera, and two objectives (1.2x and 1.5x).

Image Analysis Workstations and Software
DeltaVision Offline Deconvolution Station: a custom made system running Linux Centos 5.4 64-bit with dual Intel i7 chips, 4 GB of RAM, an Nvidia Quadro FX 3450, a 1TB drive with a data partition, and installed analysis software (DeltaVision SoftWorx, FIJI/ImageJ 64bit, OpenOffice).
FFSC 64-bit Image Analysis Station: a Dell Precision T7500 running Microsoft Windows 7 Enterprise 64-bit with dual quad-core Xeon 5590 chips, 48 GB of RAM, a Radeon 4890 graphics card, an SSD for the OS and dual 1TB SATA drives in RAID 0, and installed analysis software (Imaris 7.6, MetaMorph Premier, MATLAB, FIJI/ImageJ 64bit, Zen lite, Adobe Photoshop and Illustrator CS5, and Microsoft Office 2013).
LSRC 64-bit Image Analysis Station: a Dell Precision T7600 running Microsoft Windows 7 Pro 64-bit with an eight-core Xeon E5-2687 chip, 32 GB of RAM, an AMD FirePro W5000 graphics card, a boot drive for the OS and a 3.6TB data drive in RAID 0, and installed analysis software (Huygens SVI, MetaMorph Basic, Volocity 5, FIJI/ImageJ 64bit, and Microsoft Office 2013).
LSRC Image Analysis Station: a Dell Optiplex 990 running Microsoft Windows 7 SP1 64-bit with an Intel i7 2600 chip, 16 GB of RAM, an AMD Radeon 6670 graphics card, a boot drive for the OS and a 1TB data drive, and installed analysis software (MetaMorph Premier, Leica LAS AF offline, FIJI/ImageJ 64bit, Zen lite, Adobe Photoshop CS2, and Microsoft Office 2010).
Nanaline Duke Image Analysis Station: a Dell Precision T3500 running Microsoft Windows 7 Pro 64-bit with an Xeon W3550 chip, 24 GB of RAM, an Nvidia Quadro 4000 graphics card, a boot drive for the OS and a 2TB data drive, and installed analysis software (Imaris 8.0, Volocity 6, MetaMorph Premier, MATLAB, FIJI/ImageJ 64bit, Zeiss Zen 2010 offline, Leica LAS AF SP8 offline, and Microsoft Office 2010).
Research Park 2 Image Analysis Station: a Dell computer running Microsoft Windows 7 Pro 64-bit with an Intel i7 920 chip, 8 GB of RAM, an Nvidia GeForce GTX 260 graphics card, a 1TB drive with a data partition, and installed analysis software (MetaMorph Premier, FIJI/ImageJ 64bit, Zeiss Zen Lite, and Microsoft Office 2007).