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Electron Microscopy Sciences

EMS Academy Applications

Biological TEM Processing Workshoparrow13Biological TEM Workshop: A Complete Picture

Theory and practical preparation of buffers, fixatives, and other solutions required for chemical processing of biological samples for TEM. Sign Up

Examples of the endless possibilities in the field of Microscopy

Bone Marrow   Bone Marrow
Bone Marrow: Transmission electron microscope image of a thin section cut through an area of bone marrow area near the cartilage/bone interface in a mouse kneecap. Image shows small opening in the thin endothelium of the vascular sinus wall, where a blood cell is crossing the thin vascular sinus wall and into the sinus lumen.
Louisa Howard, Dartmouth College.
Pancreas   Pancreas
Pancreas: Transmission electron microscope image of a thin section cut through the pancreas (mouse). Image shows a cross section through part of a nerve cell and part of a capillary within the pancreatic tissue. The capillary lining consists of long, thin endothelial cells, connected by tight junctions. The image shows a fenestration in this endothelial cell. Basal lamina is present at the edges of the acinar cells.
Louisa Howard, Dartmouth College.
region in the Drosophila germarium   synaptonemal complex in nuclei of Drosophila germarium cell
Transmission electron microscope image of a region in the Drosophila germarium. Specific cells in the germarium contain synaptonemal complex in their nuclei.   Transmission electron microscope image of synaptonemal complex in nuclei of Drosophila germarium cell.
Louisa Howard, Dartmouth College.
Zebra Fish muscle   Zika Virus
Transmission electron micrograph of Zebra Fish muscle.
Specimen courtesy of Dr J Leslie.
  Transmission Electron micrograph of Zika Virus. Virus particles are 40 nm in diameter, with an outr envelope and an inner dense core.
Courtesy of Cynthia Goldsmith, CDC.

Targeted Participants

Individuals who are, or will be, responsible for chemical processing and sectioning of biological samples for TEM examination.

Facility

The EMS Microscopy Academy
Located in Hatfield, Pennsylvania, the Academy provides electron microscopy classes, workshops and training sessions for all fields of microscopy, including materials science and biological science.

Details

Tuesday - Thursday
November 7 - 9, 2017
8:00 a.m. - 4:30 p.m.
Hatfield, Pennsylvania, USA

Scope of Class

The proper chemical processing of biological samples for TEM observation is essential to maintain representative morphology and ultrastructural detail. This course will cover the buffers, fixatives, dehydrants, and embedment resins most often used for EM, with their individual advantages and disadvantages discussed. The preparation of these various solutions, when necessary, will be calculated and preformed. The microwave will be utilized for all steps except polymerization. The need for specialized protocols when using specific tissues, such as myleinated nerve which requires extended infiltration, will be discussed. Epoxy (Embed 812) and acrylic (LR White) resins will be available.

The TEM's ability to provide sub-nanometer resolution is dependent ultimately on sample thickness, typically 60 nm or less. To obtain sections of this dimension requires specialized equipment, ultramicrotomes, high quality diamond knives, and a skilled technician. The process of trimming, thick sectioning for OLM observation, thin sectioning, section retrevial, and section assessment will be the major focus of this workshop. Basic operation of the TEM will include specimen insertion, condenser and objective astigmatism correction, and critical focusing.

Format

Lecture, demonstration and hands-on practice, as well as round table discussion. Participants are encouraged to bring their own samples, if possible.

Main Curriculum

  • Theory and practical preparation of buffers, fixatives, and other solutions required for chemical processing of biological samples for TEM
  • Fixation theory and application of primary and secondary fixatives
  • Dehydration, choosing the correct one, ETOH, Acetone, or propylene oxide
  • Infiltration and choice of embedment resin, epoxy, methacrylate, or acrylic
  • Embedment and maintaining orientation if applicable
  • Block evaluation
  • Trimmimg for thick survey and thin sections
  • Practical aspects of microtomy and instrument parameters affecting section quality
  • Adjustment of clearance angle and block face:knife edge alignment
  • Thick (0.5 µm) sectioning and chromatic staining for OLM assessment
  • Thin (≤ 60 nm) sectioning and section retrevial
  • Post staining Pb and UA
  • Section assessment and troubleshooting potential artifacts such as chatter, knife marks, and tears

Instruments Available

Leica UC7 Ultramicrotome Boeckeler Autotome
EMS 9000 Microwave Vacuum Oven

Faculty

Al Coritz
Al has been working in the Electron Microscopy field for 38 years, beginning at the Yale School of Medicine and ending up on the commercial side with several key EM companies. His specialty is Cryo-techniques and Thin Film Technology: i.e. Freeze Fracture/Rotary Shadowing, High Pressure Freezing, and more. He is currently with Electron Microscopy Sciences where he has been the Technical Director for over 20 years.

Michael Kostrna
Michael was the program director of the Electron Microscopy Technician program at Madison Area Technical College and has more than 35 years in EM technical education and research experience. He has been training EM students for 29 years and has developed curricula and lab exercises for TEM, SEM, OLM, lab safety, introductory and advanced biological EM, EM, maintenance, and x-Ray microanalysis. He has worked with companies such as SC Johnson Polymer, Dow Chemicals, Io Genetics, Virent Technologies, ABS Global, NanoOnocology, and Microscopy Inovations, and in the process gained insight to the various applications of EM.

Schedule

Tuesday, November 7, 2017
8:00-8:45 Introduction of staff and participants
8:45-9:45 Buffers and fixatives used in EM
9:45-10:00 Coffee break
10:00-11:00 Dehydrants and Resins
11:00-12:00 Prepare tissues for EM processing
12:00-1:00 Provided Lunch
1:00-4:30 Divide into groups and perform station activity
     - Mixing solutions, trimming for thick and thin sectioning
     - Processing tissue
     - Sectioning, thick and thin, staining, and glass knife making
     - TEM basic operation
6:30 Hosted dinner

Wednesday, November 8, 2017
8:00-8:30 Discussion of previous day's activities
8:30-12:00 Divide into groups and perform next scheduled group activity
12:00-1:00 Provided Lunch
1:00-4:30 Divide into groups and perform next scheduled group activity

Thursday, November 9, 2017
8:00-8:30 Roundtable discussion of previous day's activities
8:30-12:00 Divide into groups and perform final group activity
12:00-1:00 Provided Lunch
1:00-1:30 Workshop assessment
1:30-4:30 Extra work on individual stations

Lodging

Participants are responsible for making their own hotel reservations. The following hotel has been designated as the host hotel:

Homewood Suites
1200 Pennbrook Parkway
Lansdale, PA 19446
Phone: 215-362-6400

The special rates are $119.00 per night (plus tax) which includes a hot breakfast and a light dinner in the evening.

Please make your reservations and mention you are participating in the EMS Workshop.
GROUP CODE: EMS WORKSHOP

Everyone should plan to arrive the evening of November 6th.

Enrollment Note

Registration will be limited to a maximum of 15 participants.
EMS will provide samples to those who prefer not to bring their own.

Registration Fee $995.00

Includes

A workshop syllabus, all supplies, reagents and solutions, lunches, coffee, tea, and dinner on the first evening of the workshop.

Online Registration

Add your registration to your cart using the links below to use our secure checkout and pay by credit card.

During checkout please indicate:

  • The name of the course
  • Will you bring your own specimens? Yes / No
  • What samples you are bringing and are most interested in?

Checkout Instructions
If only registering for a class, select any method of shipment to proceed, as shipping charges will not apply.

If ordering products in addition to registration, select desired shipping method.

Regular Tuition Registration: $995.00 Materials Ultramicrotomy Course

Pay by Check

Make payable to EMS and reference "Biological TEM Workshop".

Return your registration to:
Stacie Kirsch
1560 Industry Road
Hatfield, PA 19440 USA

Please contact Stacie Kirsch for more information.
Phone: 215-412-8400 Fax: 215-412-8450
E-Mail: stacie@ems-secure.com