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University of Kansas, Lawrence
1200 Sunnyside Avenue
1043 Haworth Hall
Lawrence, KS 66045United States of Americahttps://mai.ku.edu/
Eduardo Rosa-Molinar/ Ph.D.
Last Updated: 07/06/2022
The Microscopy and Analytical Imaging Research Resource Core Laboratory (MAI; also known as
Microscopy and Analytical Imaging Shared Resource [MAISR]), is a 2,450 ft2 core research resource core laboratory in Room 1043 in Haworth Hall at the University of Kansas Lawrence campus. The MAI (https://mai.ku.edu/) is a fee-for-service center. The imaging resources within the MAI are available to all University of Kansas, University of Kansas Medical Center, and University of Kansas Cancer Center students, staff, and faculty, students, staff, and faculty within and outside the state, and industry collaborators working in the life sciences and material sciences for nominal fees.
The MAI resource scientists (https://mai.ku.edu/lab-staff) provides training for undergraduate, graduate students, postdoctoral researchers, faculty, staff, and industry collaborators as many times as is necessary to master a given instrument and/or technique.
The MAI is directed by Eduardo Rosa- Molinar, Ph.D., Professor of Pharmacology and Toxicology, Bicampus Neuroscience Graduate Program, Anatomy and Cell Biology, and Bioengineering.
Services are offerred outside of University of Kansas, Lawrence
Consulting is offerred outside of University of Kansas, Lawrence
Hitachi 8100 Transmission Electron Microscope
Hitachi H-8100 TEM is a research grade scanning/transmission electron microscope (STEM) with analytical and high resolution imaging capabilities. Systematic theoretical instruction combined with practical hands-on training is the best way to thoroughly understand the utility and capabilities of this complex and delicate instrument. These instructions are meant only as a guide to the basic, daily alignments and operations of the H-8100, i.e. general imaging and diffraction. It is by no means a comprehensive guide to all operations of the microscope including ultra-high resolution imaging, SEM, EDX, cryo, and the various diffraction techniques. These advanced, specialized techniques will be dealt with on an as-need basis after mastery of the basic operations has been demonstrated. [Product Link]
Hitachi S4700 Field Emission Scanning Electron Microscope
Hitachi S-4700 is a field emission scanning electron microscope (FE-SEM). More than a standard SEM, the S-4700, under optimal conditions, can magnify images upwards of 500,000 times and resolve features to 2 nanometers. In addition, a cathodoluminescence detector, backscatter electron detector and energy dispersive x-ray spectrographic detector accompany the microscope. [Product Link]
Olympus IX 81 Inverted Fluorescence Automated Live cell Microscope
Olympus IX81 Inverted Fluorescense Phase Contrast Live cell Inverted microscope. [Product Link]
Leica TCS SP2 Confocal Laser Scanning Microscope
Leica TCS SP2 is a spectral confocal laser scanning microscope with 405 UV laser 3CHDIC digital emission 400-800nm range designed to acquire high spatial resolution images of fluorescently labeled materials and for analysis of these images. The confocal principle utilizes a pinhole (confocal aperture) to eliminate out-of-focus light from fluorescently labeled specimens (i.e., provide â€œoptical sectioningâ€?). Lasers provide intense, point illumination that is scanned over the preparation and the fluorescence at each point is quantified and used to construct a representation of the object brightness (an image). This method provides high resolution for the x and y planes as well as vertically (z plane). The SP2 is an advanced confocal system with nine laser excitation lines spanning the spectrum from UV to near IR. The following laser lines are available: 405nm, 458, 476, 488, 496, 514, 543, 594, and 633, all fiber coupled and controlled by AOTFs. [Product Link]
Leica EM FC6 UC6b Ultramicrotome
Leica Ultracut EM UC6 is an advanced ultramicrotome that features a Touch Screen Control Panel or a Keypad Control Panel. Both options enable fast and safer alignment of knife and specimen, and the programmable knife and cutting movements make trimming easy. With Leica's AutoTrim, you can automatically trim a specimen to a predetermined level in the block face, best for morphometric studies. Three dependent, built-in, brightness- controlled LED light sources provide outstanding illumination for top light, back light, and specimen transillumination. [Product Link]
Major equipment available:
The MAI houses five light microscopes. These include:
1. 3i/Olympus (inverted) epifluorescence optical microscope, Olympus IX 81 inverted microscope with ZDC, Prior H117 with Proscan III stage, Andor Zyla 4.2 CMOS camera, dual excitation sources, a
Lambda LS Xenon and a white LED coupled to a Sutter Lambda 10-3 filter wheel. Olympus.
– 10X 0.3NA UplanFL N, 20X 0.5NA UPlanFL N, 20X 0.75NA, 40X 0.6NA LUCPlanFL N, 40X 0.95NA UPlanSApo, 60X 0.7NA LUCPlanFL N, UPlanApo 60X 0.9NA, 100X 1.4NA UPlanSApo and 150X 1.45NA UApo N. Phase imaging available with 10X 0.3NA UplanFL N and 40X 0.6NA LUCPlanFL N, DIC imaging available with 20X 0.75NA, 40X 0.95NA UPlanSApo, and UPlanApo
Images acquired with SlideBook 6.0 using a workstation equipped with an Intel i7-6800k
processor, 32GB RAM, NVIDIA Quadro K620 graphics card, two- 256GB SSD RAID, one-2TB SATA HDD, running Windows 10 64 bit.
2. 3i/Olympus (inverted) spinning disk confocal microscope, Olympus IX-81 inverted microscope, Prior H117 with Proscan II stage, Hamamatsu Flash 4.0 v1 CMOS camera, Sutter Lambda 10-3 filter wheel, Yokagawa CSU10 spinning disk.
Olympus Objectives – 10X 0.3NA UplanFL N, 20X 0.75NA, 40X0.95NA UPlanSApo, 40X 1.3NA UPlanFL N, 60X 0.7NA LUCPlanFL N, 60X 1.42NA PlanApo N,100X 1.4NA UPlanSApo, 150X 1.45NA UApo N, CUBE 405nm laser, Sapphire 488nm laser,CrystaLaser 561 nm laser, CUBE 640nm laser, as well as the option of using a 7-line laser diode illuminator. Images acquired with SlideBook 6.0 using a workstation equipped with an Intel Xeon W3520 processor, 12GB RAM, NVIDIA Quadro FX 3500 graphics card, two-512GB SATA HDD RAID, running Windows 7 64bit.
3. Leica TCS SPE Confocal Laser Scanning Microscope (cLSM), Leica DM6-Q upright microscope, programmable piezo driven automated XYZ stage, 12-bit spectral PMT detector.
Leica Objectives –
10X 0.3NA Apo, 40X 1.15NA Apo,
Olympus Objectives - 10X 0.3NA UplanFL N, 20X 0.75NA, 40X 0.95NA UPlanSApo, 40X 1.3NA UPlanFL N, 60X 0.7NA LUCPlanFL N, 60X 1.42NA PlanApo N,100X 1.4NA UPlanSApo, 150X 1.45NA UApo N,
equipped with 405nm laser, 488nm laser, 561nm
laser, 637nm laser Images acquired with Leica LAS X using a workstation equipped with Intel i7-6800k
processor, 32GB RAM, NVIDIA Quadro K620 graphics card, two-256GB SSD RAID, one-2TB SATA
HDD, running Windows 10 64 bit.
4. Leica DM750 compound bright field upright microscope with a EXLite™ LED illumination, color
white light, a ICC50 W camera (5-megapixels) capable of connect live-stream HD to smartphones and tablets (IOS and Android), and with integrated SD card slot. Leica Objectives – 4X 0.10NA HI Plan, 10X 0.25NA HI Plan, 40X 0.65NA HI Plan and 100X 1.25 HI PLAN/Oil. Images acquired with Leica LAS CORE Imaging Software in a workstation with Intel i7-2600k processor, Intel HD Graphics card, 16 GB RAM and Windows 10 64bit.
5. The MAI houses a multi-photon excitation (MPE) microscope designed and built by Dr. Rosa-Molinar and his research group through funding from the Department of Pharmacology and Toxicology and BiCampus Neuroscience Graduate Program in collaboration with Intelligent Imaging Innovations, Inc. (3i).
This MPE microscope has the ability not only to acquire images deeper through living cells and tissue, but also integrates multiple quantitative capabilities such as, fluorescence lifetime imaging (FLIM) and time-resolved fluorescence anisotropy (polarization), to better understand physicochemical dynamics of cells, such as protein-protein interaction, protein and ligand interaction, molecular pathway analysis, and
The Zeiss Axio Examiner.Z1 is a fully motorized fixed stage upright two-photon excitation microscope workstation with a Vector™ scan head with dual galvo scanning, mBeam™ remote beam expansion, pockels cell modulator, beam delivery optics and shielding, fully integrated dual six-core computer workstation with 32GB RAM, SlideBookTM software, installation, and on-site training, four GaAsP PMTs with automatic overload and computer-controlled reset, complete filter set for multiphoton including dichroics and emission filters for simultaneous multi-color imaging (Examiner slider: 750 edge; Nosepiece input: 680 SP; Nosepiece cube: 445/45, 495 edge, 525/40; TFP/YFP: 520 LP edge, 494/34, 542/27; TFP/YFP Wide: 520 LP edge, 488/50, 550/49; Cactus input: 720 SP: Cactus cube, red/far red 617/73, 660 edge, 692/40), Additional filter cube for polarized light imaging (Cactus cube, polarized light: 525/40, s polarizer, 529/23 polarizing bandpass, p polarizer), Rapid 90° switching of the polarization of the excitation beam, Piezo objective focusing collar with a 400μm range for high-speed
volumetric imaging, Microscope filter cube for GFP for viewing epifluorescence by eye, Motorized
ZDeck system with ProScan XY stage, ProScan III controller, joystick, universal holder, solid aluminum plate and pillar spacers for Zeiss Axio Examiner.Z1, X-Cite 120LED mini white light LED system with direct coupler for Zeiss Axio Examiner.Z1, W Plan-Apochromat 20x/1.0NA water dipping objective for Zeiss Axio Examiner.Z1; working distance 2.4mm, extended IR transmission (at least 60% at 1300 nm),
48" x 60" x 4.5" aluminum optical (breadboard top) vibration-isolation table, Fully interlocked laser
safety enclosure. The two-photon excitation microscope workstation is equipped with the Chameleon Discovery, a versatile, hands-free, and fully integrated ultra-fast femtosecond laser system with a complete tuning range of 680-1300 nm. The Chameleon Discovery laser has a second output that is a fixed 1040 line, which has been combined with the tunable line to accommodate dual 2Pexcitation. One workstation equipped with an Intel Zeon E5-2620 processor, 64GB RAM, NVIDIA Quadro M2000 graphics card, one-512 SATA HDD, running Windows 10 64 bit.
MAI also houses two electron microscopes. These include:
1. Hitachi H-8100 lanthanum hexaboride (LaB6) Transmission Electron Microscope (TEM) is housed in room 1043E of the MAI. The Hitachi H8100 LaB6 TEM is equipped with an assortment of single and
double tilt, single-specimen, beryllium room temperature holders and can operate at 75 kV, 100 kV, 150kV, 175 kV, and 200 kV (75 to 200 kV), making it possible to observe biological and non-biological materials of a wide range of thickness. The Hitachi H8100 LaB6 TEM is also equipped with a Advanced Microscopy Techniques (AMT) BioSprint high speed 16 megapixel (MP) low-mount CCD camera which provides high resolution digital capture of publication-quality images. The AMT BioSprint 16 MP low-mount CCD camera is controlled by a DELL Optiplex 7070, 64 bit mini tower workstation with Intel Core i7-9700 (8 Cores/12MB/8T/up to 4.7GHz/65W), 32GB 2x16GB DDR4 2666MHz UDIMM Non-ECC, AMD Radeon R5 430, 2GB, FH, 1TB Solid State SD Hard Disk Drive, 3.5" 1TB 7200rpm SATA Hard Disk Drive, Samsung 28-Inch UE570 UHD monitor, DVD-ROM Drive, and 8x DVD+/-RW 9.5mm Optical Disk Drive, running Windows 10 Professional 64bit. The Hitachi H8100 LaB6 TEM uses hollow-cone beam illumination, a beam illumination method by which a cone shaped electron beam is produced to perform diffraction. In hollow-cone beam illumination, the beam is tilted to a certain angle against the optical axis by the first-stage deflection coils and is tilted back by the second-stage deflection coils to illuminate the same position on the specimen, then, the beam is rotated
with respect to the optical axis keeping the same illumination position. Hollow-cone beam illumination is used to eliminate diffraction contrast in bright- and dark-field images and to observe the symmetries produced only by higher-order Laue zone (HOLZ) intensity bands without strong intensity due to zeroth-order Laue zone (ZOLZ) intensity band interaction in the convergent beam electron diffraction
(CBED) pattern, composed of non-overlapping disks (Kossel-Möllenstedt [K-M] pattern) for small
convergence angles and over-lapping disks (Kossel [K] pattern) for large convergence angles.
2. Hitachi S-4700 Type II Cold Field Emission Scanning Electron Microscope (CFE-SEM) is housed in room 1043F of the MAI. The Hitachi S-4700 Type II CFE-SEM is equipped with a new generation cold
cathode field emission gun, computer-controlled flashing, extraction and accelerating voltage, Mild
flashing function, Anode heater, Accelerating voltage 500V to 30kV, Landing voltage 0.01 - 2 kV (with deceleration), Magnification 20x to 1,000,000x (referenced to 4”× 5” photo), SE resolution 0.8 nm (15 kV), 1.1 nm (1kV landing voltage), Optical Modes, Normal probe current, optimized for high resolution imaging, High probe current, increased beam current, without significant loss of resolution, for analytical techniques, Ultra high resolution, optimized for short working distances, High resolution,
optimized for long working distances, Triple Secondary Electron Detector (SED) configuration, with ExB in-column filtering, Lower SED for standard topographic imaging, Upper through-the-Lens (TTL) SED for high resolution imaging, Top TTL SED, for high resolution imaging with adjustable high pass filter function, In-lens collection system for imaging of low and/or high angle Backscattered Electron (BSE)signals, SE/BSE signal selection and mixing for TTL detectors, YAG Backscattered Electron (BSE) detector, Electromagnetic beam blanker, with automated blanking when image is frozen, 5-position objective lens aperture with built-in anticontamination heater, 6” Diameter Specimen Exchange airlock, with two touch holder exchange, 5-axis automated stage with selectable speed, Full coverage of up to a 150mm diameter sample, Specimen navigation via Rapid Image Shift Movement (RISM) and Image Navigator with optical image, Built-in specimen anti contamination device, Energy saving (ECO)
mode: 0.3kVA power consumption, One workstation equipped with an Intel Xeon E3-V3, 4GB RAM, NVIDIA Quadro K620 graphics card, one-1TB SATA HDD, running Windows 7 32 bit.
MAI houses in the KU School of Engineering's Learned Engineering Expansion Phase 2 (LEEP-2) building, room G442, houses:
1. PHI 5000 VersaProbeII XPS/AES (Physical Electronics Inc., USA) ultrahigh vacuum instrument is a raster scanned, micro focused, x-ray beam. A 20 kV electron gun, with a LaB6 emitter, is used to
create a raster scanned electron beam that is focused onto the Al anode of the x-ray source in order to create a monochromated x-rays. An ellipsoidal shaped quartz crystal monochromator collects, filters, and refocuses the x-rays onto the sample surface to scan and acquire the spectrum. VersaProbeII is equipped with dual beam charge neutralizer, sputter depth profiling (5V to 5000, Angle resolved spectroscopy, chemical state imaging, elemental composition and chemical mapping with SXI (Secondary X-ray Imaging) and Scanning Electron Microscopy (SEM; using secondary electrons), five axis automated sample manipulator with extra sample prep chamber capable for heating, cooling and reaction under mixture of different gases. The X-ray beam size for XPS ranges from 9µ to 200µ and AES is scanned at 100nm. The instrument is driven by SmartSoft software and analysis of thedata is acquired by Multipak Software tools developed by Ph ysical Electronics using a workstation equipped with a Xeon E5-1603, 4GB RAM, NVIDIA Quadro 410 graphics card, two-512GB SATA HDD RAID, running Windows 7 32 bit.
Other resources available:
The MAI contains two offline workstations with Wacom Cintiq 22 pen display. Each computer is equipped with an Intel i7-5930K processor, 128GB RAM, AMD Radeon R9 390 graphics card, one-512 SSD, one-6TB SATA HDD, running Windows 10 64 bit, one workstation equipped with an Intel i7-5930K processor, 128GB RAM, AMD Radeon R9 390 graphics card, one-512 SSD, one-6TB SATA HDD, running Windows 7 64 bit, HTC VIVE VR Headset, one workstation equipped with an AMD FX 6350 processor, 32GB RAM, AMD Radeon HD 7700 graphics card, one-128GB SSD, one-1TB
SATA HDD, running Windows 7 64 bit, one workstation equipped with an Intel Xeon 6-core
processor, 64GB RAM, ATI Radeon HD 5870 graphics card, one- 1.5TB SSD RAID, one-2TB
SATA, running MAC OS 10.9.5.
The MAI staff workstations include: two laptop workstations equipped with an Intel i7-6700HQ processor, 64GB RAM, NVIDIA Quadro M2000M, one-512GB SSD, running Windows 10 64 bit, one laptop workstation equipped with an Intel i7 4810MQ, 32GB RAM, NVIDIA Quadro K5100M graphics card, one-512GB SSD, one-1TB SATA HDD, running Windows 10 64 bit, one workstation equipped with Intel Xeon W3520 processor, 12GB RAM, AMD FirePro V3900 graphics card, one-512GB SATA HDD, running Windows 10 64 bit.
To support cell culture, cellular and tissue imaging activities we have at our disposal within the MAI
(Haworth Hall 1043) and within my laboratory (Haworth Hall 1036; 3015; 3015A) the following
1. Nikon Eclipse TE2000-E Inverted Microscope (n=1)
2. Nikon Eclipse TE2000-U Inverted Microscope (n=1)
3. Nikon SMZ 445 Stereo Zoom Microscopes (n=2)
4. Nikon Eclipse 80i Upright Microscope (n=1)
5. VWR Incubating Orbital Shaker (n=1)
6. VWR unstirred water bath to 60° C (n=2)
7. Three CO2 incubators: (1) Barnstead/Lab-Line, Model 314, (2) Shel-Lab, Sheldon Manufacturing,
Inc., Model SCO6AD, and (3) VWR CO2 incubator with 2 shaker shelves and three electric outlets
inside, multi-position stirrer and hypoxia chamber
8. Class II, Type 2 Purifier Biological Safety Cabinet, Labconco (n=2)
9. PELCO BioWave® Pro+ Microwave Processing System (n=1)
10. Leica EM UC6 ultramicrotome (n=1) with 35° 45° (n=1) diamond knives
11. Sorvall MT-2 (n=2) and MT-2B (n=5) ultramicrotomes with 35° (n=7), 45° (n=7), Gumbo Histo (n=1), and oscillating (n=1) diamond knives
12. Sorvall TC-2 Tissue Slicer (n=1)
13. Sorvall GKM Knifemaker (n=2)
14. Leica EM KMR3 Knifemaker (n=1)
15. American Optical Spencer 860 Sliding Block Microtome (microtome knife blade 185 mm 7.25 inch) with Cryo-Histomat MK-2 controller, 75mm x 80mm stage freezing stage, closed circuit cooling
system and (n=1)
16. American Optical Spencer 820 Rotary Microtome with AO 820 Blade Holder #814, High
Profile Disposable Blades, LAB-TEK AO820 cassette vise jaws with floatation bath workstation(n=1)
17. Quorum Technologies carbon/metal evaporator/plasma cleaning (n=1)
18. Quorum Technologies critical point dryer (n=1)
19. Cryopress Slam Freezer (n=1)
20. FEI Vitrobot (n=1)
21. Cold storage facilities consisting of a liquid nitrogen freezer (n=1); cryogenic storage Dewars (n=6);
ultra-low upright freezer to -85° C (n=1), laboratory freezers to -15° C (n=4) and laboratory
refrigerators to 4° C (n=4)
Publications associated with this facility (Click To View):
1.) Reynolds KA, Rosa-Molinar E, Ward RE, Zhang H, Urbanowicz BR, Settles AM (2022 Feb 5). Accelerating Biological Insight for Understudied Genes. Integrative and comparative biology, 61(6), 2233-2243. . ID: 33970251.
2.) Kopec BM, Kiptoo P, Zhao L, Rosa-Molinar E, Siahaan TJ (2020 Feb 3). Noninvasive Brain Delivery and Efficacy of BDNF to Stimulate Neuroregeneration and Suppression of Disease Relapse in EAE Mice. Molecular pharmaceutics, 17(2), 404-416. doi: 10.1021/acs.molpharmaceut.9b00644. ID: 31846344.
3.) Kopec BM, Zhao L, Rosa-Molinar E, Siahaan TJ (2020 Feb). Non-invasive Brain Delivery and Efficacy of BDNF in APP/PS1 Transgenic Mice as a Model of Alzheimer Medical research archives, 8(2), . pii: 2043. doi: 10.18103/mra.v8i2.2043. ID: 32551362.
4.) Song JY, Larson NR, Thati S, Torres-Vazquez I, Martinez-Rivera N, Subelzu NJ, Leon MA, Rosa-Molinar E, Schöneich C, Forrest ML, Middaugh CR, Berkland CJ (2019 Jan 10). Glatiramer acetate persists at the injection site and draining lymph nodes via electrostatically-induced aggregation. Journal of controlled release : official journal of the Controlled Release Society, 293(), 36-47. . ID: 30414463.
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University of Kansas Microscopy and Analytical Imaging Research Resource Core Laboratory Core Facility (RRID:SCR_021801)
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