The Imaging Facility Newsletter

Inside this issue:

Featured

Microscope 2

Zeiss 710 Update 3

Celebrate

Biomedical Art 4

IF Updates 5

IF Survey Results 6

Recent Publications 7

Contact Us 10

I am a 2nd year Masters student in Dr. Pistilli’s lab in the Exercise

Physiology Department. We study the effects of cancer cachexia on

skeletal muscle wasting and function. We recently classified a murine

-derived mammary tumor cell line as having the ability to form

tumors in vivo and induce cachexia within 4 weeks post injection.

We monitored tumor status weekly using both optical and

ultrasound imaging, and we performed physiological assessment of

muscle function.

Large-stitch images of immunostained muscle cross sections were

acquired using the Nikon TE2000 Sweptfield microscope. These

images were then analyzed using the NIS Elements General Analysis

program to automatically determine muscle fiber area. We showed

severe fatigue at the 4 week time point for our mice, highlighting

both skeletal muscle wasting and dysfunction. Building upon these

results, we repeated these experiments in our transgenic model of

IL-15 skeletal muscle overexpression where we observed

preservation in function and fiber area. We are now continuing to

characterize the effectiveness of IL-15 in the attenuation of cancer

cachexia.

Volume 5, Issue 2

August 2016

Featured Researcher: Joseph Bohlen

Large-stitch image of a mouse muscle cross section (left). Muscles were labeled with laminin

and DAPI, and using the Nikon Sweptfield microscope, 25 images at 20X were captured and

stitched to show the entire section. Since the individual images were obtained with a 20X

objective lens, zooming in to an area of the muscle does not result in loss of resolution.

IVIS optical imaging (left) was used to non-invasively monitor tumor growth over time. A

series of 2D ultrasound images (middle) were collected and reconstructed into a 3D structure

(right) for accurate tumor volume measurements.

Page 2

Lightning-Fast

Brightfield

Scanning

3D Virtual

Slides

Multi-Channel Fluorescent

Slides

New Equipment in the Imaging Facility!

6 Position

Slide Holder

for High

Throughput

The Microscope Imaging Facility has recently installed a new Olympus VS120 Slide Scanner

to replace the old Olympus Histology

Microscope.

This system can image entire tissue sections,

either in brightfield or fluorescence.

This system was purchased with funding from the INBRE

grant 5P20GM103434 and the WVU HSC Office of

Research & Graduate Education. Please remember to

acknowledge this support in your publications.

Olympus VS120

Slide Scanner

The system is equipped with 5 objectives for

2x - 40x magnification.

This scanner is super-duper fast and easy to

use.

Please contact Mandy Ammer for a demo or

to setup training.

Page 3

Zeiss 710 Confocal with Airyscan

There are two new objectives on the Zeiss 710 confocal.

40X Water 63X Oil

Airyscan Technology on the Zeiss 710 Confocal

Airyscan imaging yields a 1.7x enhancement in lateral resolution.

The system now has two super-resolution quality objectives.

Beam alignment for the detector takes place in just a few

minutes.

Please contact Mandy Ammer in the Imaging Facility to schedule

your training session today!

Please Handle with Care!

These are very expensive objectives.

Celebrate Biomedical Art Page 4

The National Institutes of Health Life: Magnified exhibition has turned

life into art. This gift, displayed in the Pylons area and library at the

Health Sciences Center, will enlighten and educate for years to come.

Join the celebration of this exhibit at a reception in the Pylons lobby

Wednesday, August 24, 2016, 3:30 - 5:30 p.m.

In addition, the NIH has contacted us looking for more breath-taking

images to add to their collection. If you have images you’d be willing

to share, please let us know and we’ll help pass them along.

The IF would like to wish Emily Ellis the best of luck in her new

position in the laboratory of Dr. Timothy Eubank.

We will miss you!

newsletter

Page 5

New Server For Storing Your Awesome Images The Imaging Facility Users now have space on a brand

new server that will provide tons of room for data

storage. All of your data in your AMIF/MIF User Folders

was successfully transferred to the new location, and it

is ready to use.

You should now have a shortcut named "Imaging Folders" on your desktop that will

take you to the new server location; you should also see this location mapped to your

P: drive. If you don't see these links, try logging out and back in and/or restarting your

computer. If that doesn't work, let us know and we'll figure out how to make those

connections.

\\hs-mbrcc\mbrcc\RESLAB\Imaging (for PCs)

smb://hs-mbrcc/mbrcc/reslab/imaging/ (for Macs)

Service Rate/Hr

AMIF IVIS Lumina II $30.00

AMIF Vevo 2100 $38.50

AMIF XenX $20.00

AMIF SkyScan 1272 $16.50

AMIF Procedures Room $5.50

Assistance/Procedures/Training $30.00

The Core Resources Management Committee (CRMC) has recently completed the

annual review of the Imaging Facilities, and the following rate schedule was approved

effective October 1, 2016.

Imaging Facility Rates Starting Oct 1

Service Rate/Hr

Assistance/Training $30.00

MIF Nikon Live Cell $15.00

MIF Nikon Sweptfield (<4 hours) $15.00

MIF Nikon Sweptfield (> 4 hours) $10.00

MIF NLOM 2-Photon $15.00

MIF Olympus MVX10 $0.00

MIF Olympus Slide Scanner $10.00

MIF Zeiss 710 Confocal (<4 hours) $25.00

MIF Zeiss 710 Confocal (>4 hours) $18.50

MIF Zeiss Fluorescent $15.00

MIF Zeiss PALM Laser Microdissection $15.00

MIF Zeiss Violet Confocal $30.00

Workstations $5.00

Microscope Imaging Facility Animal Models & Imaging Facility

Imaging Facility Surveys Page 6

Thanks to everyone who has taken our Imaging Facility Surveys over

the past year! Your feedback is really important for us to learn what

we’re doing well and what we need to improve to best support your

research.

Here’s a

summary of

what you

told us.

Your feedback is also critical in identifying new equipment needs.

Here are a couple of requests and an update on what we’re doing...

*Two-photon microscope with live animal imaging capabilities is needed.

The imaging facility is working with Nikon to bring in a two-photon

microscope for a demo, hopefully later this fall. Please let us know

ASAP if you’d like to participate. We will need to get ACUC protocols

submitted soon in order to make the most of this demonstration.

*Super resolution imaging system is needed.

We hosted a demo of the Nikon A1R confocal / N-SIM super-

resolution imaging system this spring. With your help, we had a very

successful demo and were able to submit an instrumentation grant to

fund the purchase of this system. Our application will be reviewed in

November - we’ll keep you posted.

For training and assistance from the AMIF, please

keep in mind that Sarah’s regular schedule is Monday,

Tuesday and Thursday 7:30 a.m. - 5:30 p.m. The

calendar can fill up quickly, so please schedule in

advance to make sure we can accommodate your

imaging needs.

Survey Results (Scale of 1 to 5) AMIF MIF

Availability of facility & equipment 4.7 4.5

Quality of results/data 4.4 4.5

Availability of staff 4.7 4.7

Knowledge and expertise of staff 4.3 4.8

Quality of consultation 4.6 4.8

Cost of service 4.0 3.8

Overall experience 4.7 4.7 Take the

AMIF

Survey

Take the

MIF

Survey

https://goo.gl/forms/

BMzvGPCw7JLKUzXW2

https://goo.gl/

forms/4CLrBVbanZpDNYCj

Page 7

Early detection of cardiac dysfunction in the type 1 diabetic heart using speckle-

tracking based strain imaging. Shepherd DL, Nichols CE, Croston TL, McLaughlin SL,

Petrone AB, Lewis SE, Thapa D, Long DM, Dick GM, Hollander JM. J Mol Cell Cardiol.

2016 Jan;90:74-83. PMID: 26654913

Cardiac and mitochondrial dysfunction following acute pulmonary exposure to moun-

taintop removal mining particulate matter. Nichols CE, Shepherd DL, Knuckles TL,

Thapa D, Stricker JC, Stapleton PA, Minarchick VC, Erdely A, Zeidler-Erdely PC, Alway

SE, Nurkiewicz TR, Hollander JM. Am J Physiol Heart Circ Physiol. 2015 Dec 15;309

(12):H2017-30. PMID: 26497962

Deficiency of isoprenylcysteine carboxyl methyltransferase (ICMT) leads to progres-

sive loss of photoreceptor function. Christiansen JR, Pendse ND, Kolandaivelu S, Ber-

go MO, Young SG, Ramamurthy V. J Neurosci. 2016 May 4;36(18):5107-14. PMID:

27147662

Grainyhead-like 2 inhibits the coactivator p300, suppressing tubulogenesis and the epi-

thelial-mesenchymal transition. Pifer PM, Farris JC, Thomas AL, Stoilov P, Denvir J,

Smith DM, Frisch SM. Mol Biol Cell. 2016 Aug 1;27(15):2479-92. PMID: 27251061

Recent Publications

Shepherd et al. Nichols et al.

Christiansen et al. Pifer et al.

Page 8

Please let us know

when you publish a manuscript with da-

ta from the MIF or AMIF so that we may acknowledge

your achievement in

our newsletter!

Salubrinal reduces oxidative stress, neuroinflammation and

impulsive-like behavior in a rodent model of traumatic brain injury.

Logsdon AF, Lucke-Wold BP, Nguyen L, Matsumoto RR, Turner RC,

Rosen CL, Huber JD. Brain Res. 2016 Apr 27. pii: S0006-8993(16)

30312-2. PMID: 27131989

Endoplasmic reticulum stress implicated in chronic traumatic

encephalopathy. Lucke-Wold BP, Turner RC, Logsdon AF, Nguyen L,

Bailes JE, Lee JM, Robson MJ, Omalu BI, Huber JD, Rosen CL.

J Neurosurg. 2016 Mar;124(3):687-702. PMID: 26381255

In Vitro expansion of bone marrow derived mesenchymal stem

cells alters DNA double strand break repair of etoposide induced

DNA damage. Hare I, Gencheva M, Evans R, Fortney J, Piktel D,

Vos JA, Howell D, Gibson LF. Stem Cells Int. 2016; 2016:8270464.

PMID: 26880992

Bcl6 modulation of acute lymphoblastic leukemia response to

chemotherapy. Slone WL, Moses BS, Hare I, Evans R, Piktel D,

Gibson LF. Oncotarget. 2016; 7(17): 23439-53. PMID: 27015556

Recent Publications Continued

Logsdon et al.

Moses et al.

Lucke-Wold et al.

Addison et al.

Hare et al.

Recent Publications Continued Page 9

Please Remember to Acknowledge Us!

KAP1 promotes proliferation and metastatic progression of breast cancer cells. Addison

JB, Koontz C, Fugett JH, Creighton CJ, Chen D, Farrugia MK, Padon RR, Voronkova MA,

McLaughlin SL, Livengood RH, Lin CC, Ruppert JM, Pugacheva EN, Ivanov AV. Cancer Res.

2015; 75(2): 344-55. PMID: 25421577.

Grainyhead-like 2 reverses the metabolic changes induced by

the oncogenic epithelial-mesenchymal transition: effects on

anoikis. Farris JC, Pifer PM, Zheng L, Gottlieb E, Denvir J,

Frisch SM. Mol Cancer Res. 2016; 2016; 27(15): 2479-92..

PMID: 27084311

Bone marrow microenvironment niche regulates miR-221/222

in acute lymphoblastic leukemia. Moses BS, Evans R, Slone WL,

Piktel D, Martinez I, Craig MD, Gibson LF. Mol Cancer Res.

2016 Jun 29. PMID: 27358112

In vivo assessment of coronary flow and cardiac function after

bolus adenosine injection in adenosine receptor knockout

mice. Teng B, Tilley SL, Ledent C, Mustafa SJ. Physiol Rep. 2016

Jun;4(11). PMID:27302991

AMIF: “Small animal imaging and image analysis were performed in the West Virginia University

Animal Models & Imaging Facility, which has been supported by the WVU Cancer Institute and

NIH grants P20 RR016440, P30 GM103488 and S10 RR026378.”

MIF: “Imaging experiments and image analysis were performed in the West Virginia University

Microscope Imaging Facility, which has been supported by the WVU Cancer Institute and NIH

grants P20 RR016440, P30 GM103488 and P20 GM103434.”

Farris et al. Slone et al.

Teng et al.

1 Medical Center Drive

Erma Byrd BMRC

West Virginia University

Morgantown WV 26506

Karen Martin

kamartin@hsc.wvu.edu

Phone: 304-293-6965

Fax: 304-293-4667

The purpose of this newsletter is to inform researchers about the AMIF and MIF.

We want all investigators, graduate students and staff to be knowledgeable about

the equipment and resources that are available. The staff are always glad to

discuss upcoming studies with investigators to best utilize the core resources

available. To learn more about our facilities, please check out our websites (to

the left) or contact us to speak directly with AMIF or MIF staff.

wvucancer.org/AMIF

Contact Us!

wvucancer.org/MIF

Sarah McLaughlin

smclaughlin@hsc.wvu.edu

Phone: 304-293-0518

Fax: 304-293-4667

Mandy Ammer

aammer@hsc.wvu.edu

Phone: 304-293-0942

Fax: 304-293-4667

Page 10 Humor to encourage your science

Picture Perfect Image Analysis using

Nikon NIS Elements Tuesday, September 27 at

1:00 PM

Erma Byrd 201