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Welcome to the East Carolina University Imaging Core Facility

Our goal is to provide comprehensive and state of the art imaging for a wide-range of biological and other disciplines on and off the East Carolina University Campus for both research and teaching.

Currently we can provide imaging in Conventional and Variable Pressure Scanning Electron Microscopy, Transmission Electron Microscopy, and Confocal Fluorescence Microscopy. Details concerning our core equipment can be found under the Core Imaging Equipment tab.

We can be found in the Howell Science Building in rooms C302-C307.

Please feel free to contact us using the information below.

Director
Dr. Anthony Capehart
252-328-6296
capehartt@ecu.edu

Lab Manager
Dr. Tom Fink
252-737-2300
finkt@ecu.edu

Our website is new and will be evolving rapidly, please check back for new additions and changes.

Major support equipment and software are listed below under five categories. Click here more information about each support item.

SEM

Bal-Tec CPD 030 Critical Point Dryer
Anatech Hummer Model 6.6 Sputter Coater (Gold/Palladium)
Anatech CEA 2.2 Carbon Evaporation Accessory for the Hummer 6.6

TEM

Leica Reichert Glass KnifeMaker II
RMC, Inc. MT6000-XL: Motorized Microprocessor Controlled Ultramicrotome
Sorval MT2: Motorized Ultramicrotome
Fisher Vacuum Oven
Lynx el Microscopy Tissue Processor
Epson Perfection V700 Photo Film Scanner for 4”x5” TEM negative scanning

Paraffin Thick Sectioning

Reichert-Jung Model 2030 BioCut Paraffin Sectioning Microtome
Leica SP 9000 Knife Sharpener
Microm Cryostat Microtome HM 550

General

Nikon Super CoolScan LS-9000 ED Film Scanner

Imaging Software

Adobe Photoshop CS4: Image Enhancement, Scale bar generation, Scientific Plate Formation
Adobe Illustrator: Graphics
Powerpoint: Oral and Poster presentations of data
Image J, NIH Image, Scion Image PC: Computer measurement of micrographs
Slidebook 4.20: In addition to control of the Cofocal Microscope this software allows for sophisticated measurements such as 3-D volume of structures captured in a z-series

To use the Quanta 200 SEM, Philips CM12 TEM, Olympus IX2-DSU Confocal Mircroscope and the Olympus BX-41 Fluorescence/Phase microscope you will sign up for use on our Sharepoint Website. If you have not been added as a user to the Microscopy Sharepoint Website, then see Tom Fink or e-mail him at finkt@ecu.edu and he will add you as a user and there are instructions on the Sharepoint website on how to sign up for core imaging equipment. Only East Carolina University students, faculty and staff can access our Sharepoint site, outside users would have to contact Tom Fink directly.

To use support equipment in the imaging core facility please contact Tom Fink.

Our Fees are an attempt to recoup a portion of the costs incurred in operating a microscopy facility. You can when appropriate save money by doing any specimen preparation in your laboratory, at your time, and with your supplies. The following FEES for ECU personnel are for those with grants in which microscopy fees have been budgeted for. If you do not have grant support, please contact us directly and we will try to accommodate your needs.

1. Fees for ECU personnel (a run involves specimens in one to six 3 ml vials): 

a. Scope Time Per Hour (Quanta 200 SEM, Philips CM12 TEM, Olympus IX2-DSUConfocal: $30.00 unassisted, $50.00 assisted (includes training) 

b. TEM Film and Developing: $3.00/picture: Includes film, developer chemicals, digital scanning 

c. SEM Specimen Preparation:

i. Fixation: $15.00/run, $30.00/run with technician 

ii.Dehydration: 

1. Critical Point Drying: $15.00/run, $45.00/run with technician

2. Hexamethyldisilizane: $10.00/run, $20.00/run with technician 

iii. Sputter Coating: $10.0/run, $20.00/run with technician 

iv. Specimen mounting: $1.00/stub

d. TEM Specimen Preparation:

i. Fixation, dehydration, infiltration, embedding: $50.00/run

ii. Sectioning: $36.00/hour if we do it, glass knife cost if you do it.

iii. Grid Staining: $5.00/grid if we do it, grid and chemical cost if you do it. 

2. Non-ECU personnel and Commercial: Please contact us for custom quotes of fees.

We follow the laboratory safety parameters of the Office of Environmental Health and Safety at East Carolina University. In general there are little or no hazards involved in using the microscopes. The chief hazards involve chemical preparation of samples for transmission electron microscopy (TEM) and conventional scanning electron microscopy (CSEM). On the main computers in C302 and C307 there is a shortcut on the desktop at the upper right corner (icon is a gold star) to a file titled Lab Safety at ECU. In that folder is the following pertinent safety information: 0_Safety at ECU (includes this document and ECU Chemical Hygiene Plan 2007), 1_Standard Operating Procedures (with focus on safety), 2_Laboratory Safety Plans, 3_Material Safety Data Sheets, and other safety information (total of 7 folders, 0-6).

Safety Links

Office of Environmental Health and Safety at East Carolina University:
http://www.ecu.edu/oehs/

Georgia Tech:
www.safety.gatech.edu

Georgia Tech also has a very nice powerpoint lecture on general chemical safety. Click on the link below and then click on the “Safety Seminar Slides” near the right bottom of that page.
http://www.mse.gatech.edu/Research/lab_safetypolicy/lab_safetypolicy.html

Tutorials

Computer based tutorials (Powerpoint, Word, Camtasia videos) can be a powerful way to learn, assist learning, and review many aspects of microscopy techniques. Our goal is to produce learning aids for most of the core imaging equipment and support equipment, and for using critical image analysis software. Please check back periodically to view the latest entries and updates. We also invite you to create your own learning tutorials that you might like to share with the ECU community. There will be different levels of tutorials from simple directions for experienced users to tutorials with many photographs and/or videos.

To access these tutorials click on one of the pull-down menus below for : 1. Scanning Electron Microscopy Tutorials, 2. Transmission Electron Microscopy Tutorials, 3. Confocal Microscopy Tutorials, and 4. General Imaging and Image Analysis Tutorials.

Scanning Electron Microscopy Tutorials

Transmission Electron Microscopy Tutorials

Confocal Microscopy Tutorials

General Imaging and Image Analysis Tutorials

Teaching

We currently teach a Scanning and a Transmission Electron Microscopy course during alternate spring semesters. These courses involve considerable hands on experience on the scopes and lead to considerable proficiency in skill of microscope usage. The goal of each course is to produce a research grade poster on a subject or technique that is ultimately publishable and/or useful to research programs in the Department of Biology and elsewhere at ECU. Current course syllabi can be viewed below (SEM Syllabus followed by the TEM Syllabus). Following the syllabi are 5 examples of Student SEM posters from April 2009.

SEM Syllabus
TEM Syllabus

Aaron Wallace's Poster
Andrew Freistaedter's Poster
Anna Wroblewska's Poster
Drew Cathey's Poster
Charles Worley's Poster

Microscopy References

SEM, TEM, and Light Microscopy

  1. Bozzola John, J. and Russell, Lonnie D. 1999. Electron Microscopy, Second Edition. Jones and Bartlett Publishers. 670 pp. 
  2. Kuo, John, editor. 2007. Electron Microscopy Methods and Protocols, Second Edition. Humana Press, Inc., 608 pp. 
  3. Chandler, Douglas E. and Roberson, Robert W. 2009. Bioimaging. Current Concepts in Light and Electron Microscopy. Jones and Bartlett Publishers, 440 pp.

SEM:

  1. Goldstein, Joseph I. et al. 1992. Scanning Electron Microscopy and X-Ray Microanalysis. A Text for Biologists, Materials Scientists, and Geologists, Second Edition. Plenum Press, 820 pp. 
  2. Goldstein, Joseph I. et al. 2003. Scanning Electron Microscopy and X-Ray Microanalysis, Third Edition. Springer, 690pp + DVD. 
  3. Echlin, P. (ed.). 2009. Handbook of Sample Preparation for Scanning Electron Microscopy and X-Ray Microanalysis. Springer, 330pp. 
  4. Schatten, H. and Pawley, J. B. (eds.). 2008. Biological Low-Voltage Scanning Electron Microscopy. Springer, 318pp.

TEM:

  1. Williams, D. B. and Carter, C. B. 2009. Transmission Electron Microscopy. A Textbook for Materials Science. Second Edition. Springer, 850pp. 
  2. Frank, J. (ed.). 2006. Electron Tomography. Methods for Three-Dimensional Visualization of Structures in the Cell. Springer, 455pp.

Confocal/Fluorescence Microscopy:

  1. Hibbs, Alan R. 2004. ConfocalMicroscopy for Biologists. Springer, 467pp. 
  2. Pawley, J. B. (ed.). 2006. Handbook of Biological Confocal Microscopy. Third Edition. Springer, 988pp. 
  3. Muller, Michiel. 2006. Introduction to Confocal Fluorescence Microscopy, Second Edition. SPIE Press, 120 pp.

Microscopy Links

Microscopy Societies:

Microscopy Society of America:
http://www.microscopy.org/

General Microscopy

The University of Arizona has very nice website with numerous links to microscopy related information of all types. Look under the Resources tab.:
http://swehsc.pharmacy.arizona.edu/exppath/resources/tutorials.php

Olympus Microscopy Resource Center:
http://www.olympusmicro.com/

General Histology Techniques/Paraffin Thick Sections:

WEB PATH: The Internet Pathology Laboratory for Medical Education, Mercer University School of Medicine, Savannah; The University of Utah, Eccles Health Sciences Library:
http://library.med.utah.edu/WebPath/webpath.html#MENU

Reflected Fluorescence Microscopy:

Filter Handbook from Chroma Technology Corp.:
http://www.chroma.com/resources/filter-handbook

Digital Image Measurement Software:

The NIH Image website is a great place to download free software for image analysis and measurement. Programs include NIH Image (for Macintosh computers), Scion Image PC (Windows), and Image J (Windows and Macintosh). There are many other programs.
http://rsb.info.nih.gov/nih-image/

The purpose of this section is to give the viewer a glimpse into the capabilities of the Imaging Core Facility. It is divided into three sections: SEM Images and Anaglyphs, TEM Images, and Confocal Images. Each segment of the gallery involves a different research project where the images are described in the context of the project or study. Studies may utilize more than one type of microscopy but will be filed under one of the three imaging microscopies mentioned above. Please also view the student posters under the Teaching Tab (Tab # 6) and the Outreach Tab (Tab # 10). The Gallery section of the website is truly in progress, please check back frequently to view new additions to the content.

SEM and Anaglyphs Gallery

Empis snoddyi Empis snoddyi silk bubblewrap

Empis snoddyi males make and carry a silk nuptial gift for attracting females (Jennifer A. Sadowski et al. 1999. Behav. Ecol. Sociobiol. 45: 161-166) (video link of male swarming). This very fragile structure would be damaged by any handling and sample preparation. It was successfully imaged in Low Vacuum mode of our Quanta 200 SEM. The surface was imaged and also the entire depth of the gift wall thereby showing that the wall is made up of variable “multigons” , kind of like variable bubble wrap.

TEM Images

none

Confocal Images

Drosophila melanogaster

Drosophila melanogaster (Fruit Fly) egg chamber. DNA appears blue while Actin appears in green. Follicle Cells (which manufacture the chorion or egg shell) surround the developing oocyte which is nourished by the large nurse cells at the base.

We relish the idea of exposing students of all ages to the capabilities and wonders of different modes of microscopy/imaging. Since September 2009 we have had some educational programs involving 4-H students of a wide range of ages and some local high-school students. Please feel free to contact us for your imaging education desires.

aphidbody aphidhead

 headcloseup aphidtail

These Aphids from a cotton leaf in the ECU Biology Greenhouse were collected at 10:30 AM 30 June 2009 and then imaged directly with no specimen preparation at all in the Low Vacuum mode of the Quanta 200 SEM at 1 Torr and room temperature. The Aphids were still alive 2.5 hours later!

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