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Electron Microscopy for Pathologists
Introduction
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Introduction to the Resident Rotation
Learning Objectives
About the TEM
Tissue Preparation
Alternative Tissue Preparation from
Paraffin Block
Ultramicrotomy
Darkroom Procedures and Digital Imaging
List of Required Cases Introduction to the Resident Rotation
EM faculty and staff train Pathology Residents during a one-month rotation
in each Resident's second year. The
rotation provides residents an opportunity to gain skills in the use of the
transmission electron microscope (TEM). Additionally, teaming with faculty mentors, they
gain knowledge and abilities in the development of ultrastructural
diagnoses. Current biopsy and autopsy materials, as well as cytological
and clinical specimens are available for their studies. Learning
Objectives
Upon
completion of the rotation, residents are able to:
-
Independently
operate the transmission electron microscope (TEM).
-
Applying departmental
protocols, independently
prepare tissue specimens applying fixation, embedding, and sectioning
procedures, as well as alternative alternative
paraffin
block procedures.
-
Discuss ultramicrotomy, including the cutting of thick and thin sections.
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Describe
darkroom procedures and digital imaging of EM negatives.
-
Recognize
ultrastructural characteristics of well-fixed normal tissues and
processing artifacts.
-
Correctly
diagnose surgical cases on file using the TEM.
The
rotation is the Residents only scheduled opportunity for hands-on
exposure to EM. Other training and practice are available to VCU/MCV faculty and
staff by appointment.
About
the Transmission Electron Microscope (TEM)
The TEM is similar in function to a
light microscope. It requires thin specimens that allow electron
penetration. The development of the light microscope took about 370 years,
while the present-day electron-utilizing instruments are the result of
less than 50 years development. The importance of the electron microscope
to biology is seen in its resolving capabilities--0.1 to 0.2
nanometer-sized molecules--over the resolving power of the light
microscope which is 0.2 micrometer-sized particles.
The electron microscope is similar in construction to a light
microscope, except that it is inverted. The electron gun acts as the
source of illumination, taking the place of the light microscope lamp. The
glass lenses are replaced by electromagnetic lenses (magnetic fields which
focus the electron beam). Since electrons cannot be seen by the human eye,
a viewing screen, coated with a material that fluoresces is used (usually
zinc sulfide crystals). The three basic lens systems--condenser,
objective, and projector lenses--function as in the light microscope.
There is a mechanical stage on which the specimen is placed.
To speak directly with someone
concerning EM training please
contact:
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