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Photos of Results Achieved with
BioWave® Pro Microwave Processing Technique
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Fig. 1A-C. Figure 1A is a micrograph of a normal sural nerve with a non-myelinated nerve (N) having secretory
vesicles (sv), microtubules (mt) and a swan cell nucleus (ScN). The insert (1B)
shows a myelinated nerve and the
arrows clearly demonstrate its periodicity. Figure 4C is a membranouse Lupus nehpritis
(RPS/ISN Class V). There
is diffuse, generalized effacement of the foot processes of the visceral epithelial
cells. Numerous regularly
disposed epimembraneous immune complex deposits are illustrated by the arrows. Both
tissues were initially fixed
in a variant of 10% NBF (Carson et al. 1972) and then processed in the microwave
for ultrastructural evaluation by
the methods of Giberson et al. (2003) and Austin (2002). Micrographs from Ronald
L. Austin, Research Associate,
Dept. of Pathology, LSU Medical Center, Shreveport, LA 71130.
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Figure 2A-G. Fig. 2A-B shows cytoplasmic iridovirus from the skin of a sturgeon. The iridovirus
is a large enveloped dsDNA virus which infects both insect and vertebrate hosts.
Fig. 2C-E demonstrates an intranuclear
baculovirus from the hepatopancreas of a crayfish from Northern California. Fig.
2C is a low magnification image of the
enveloped dsDNA virus showing the intranuclear arrangement of virus particles Fig.
2D is a higher magnification showing
both a cross-sectional and longitudinal view of the virus. Fig. 2E is a high magnification
cross-section of a number of
virus particles demonstrating the unique intranuclear membrane-bound virions. Fig. 2F-G demonstrates an endothelial cell
polyoma virus from a blood vessel in the liver of a parakeet. Polyoma virus have
a single molecule of circular dsDNA and
the particles are non-enveloped. Polyoma virions are spherical in outline and typically
45nm in diameter. Fig. 2F is a
low magnification image showing typical nuclear presentation. Fig. 2G is a high
magnification view of the virus. Infected
tissues were processed directly from 10% NBF by the microwave methods of Nordhausen
and Barr (2001) and Nordhausen et al.
(2002). Micrographs from Bob Nordhausen, Univ. of California, Davis, California
Animal Health and Food Safety Lab, School
of Veterinary Medicine, Davis, CA 95616.
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Figure 3. Micrographs from a 2008 microwave workshop held at the Center for Microscopy, San Joaquin Delta College,
Stockton, CA. Rat brain (not perfusion fixed) (1), cardiac muscle (2), kidney (3)
and liver (4) were processed from
osmium through resin polymerization for a net turnaround time of under 4 hours from
fresh tissue to the electron
microscope. Microwave techniques (Giberson, et al., 2003) make it possible to teach
and in real time demonstrate
the techniques of electron microscopy.
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Figure 4A-F. Figures A-C are of normal mouse liver benched fixed in 10% NBF for 24 hours. Figures D-F
are of normal mouse liver fixed in 10% NBF for 20 minutes utilizing microwave radiation.
All tissues were prepared for
fixation identically and cut to 2mm prior to fixation. Figures A and D are corresponding
Hematoxylin and Eosin stained
sections and figures B and E are corresponding Vimentin IHC stained sections. Figures
C and F are corresponding EM sections
demonstrating complimentary ultra-structure. Images are from Dr. Jose Galvez, Center
for Comparative Medicine and Department
of Medical Pathology, University of California, Davis, CA.
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Figure 5. Tissues formalin fixed and paraffin processed by the protocols described in Galvez et al. 2006.
Mouse mammary tumor virus induced mammary carcinoma (A, B). Note the mitotic figures
(arrows) in B. Mouse esophagus with clearly identifiable muscle striations (*) (C).
Mouse uterus stained with mouse anti-estrogen (D). (Center for Comparative
Medicine and Department of Pathology, Univ. of Calif., Davis)
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Figure 6: Confocal projection of Elodea canadensis labeled with Hoechst 33258 nucleic acid probe (blue stain) for
6 minutes in the absence (A) and presence (B) of 150W microwave radiation. Confocal
projection of Arabidopsis thaliana
root tip labeled with Hoechst 33258 nucleic acid probe after 6 minutes of 150W microwave
radiation (C). Confocal projection
of in situ hybridization patters of whole chromosome probes (red) hybridized to nuclei
of paraffin embedded rabbit skin (D).
Confocal projection of mouse kidney paraffin sections labeled with anti-Factor VIII
monoclonal antibody for 60 minutes on the
bench (E) and after 6 minutes of 150W radiation (F) (Scale for all bars = 50µm) (Table
1). (Mark Sanders, Imaging Center, College of Biological
Sciences, Univ. of Minnesota, St. Paul, MN). Reprinted with permission of Galvez
et al., Microscopy and Analysis, Nov. 2006.
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Figure 7. Retinas were fixed in 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4) overnight at 4°C. Following fixation the
tissue was rinsed 6 x 20 minutes in buffer prior to beginning antibody labeling.
The bench staining protocol required 7 days. The
labeling results were completed in an afternoon using microwave-enhanced labeling
during a workshop held at the Univ. of Minnesota
Imaging Center (Mark Sanders, Director - May 17-19, 2006). The retinas were triple-labeled for:
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Figure 8. Note the symmetrical pattern of demineralization when microwave methods are employed.
The separated piece at the top broke off during plastic embedding. From the work
of Steven P. Tinling (Tinling et al., 2004),
Otolaryngology Research Laboratory, University of California, Davis 95616.
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Figure 9. Left Image. Image on the left is an electron micrograph of an inner hair cell (IHC) and supporting cells (S)
from a Japanese macaque monkey. The lower left arrow indicates the region shown
at higher magnification in the image to the right. Bar = 3.0µm.
Right Image. In the supporting cells next to the inner hair cell, the rough endoplasmic
reticulum and microtubules are well-preserved. Bar = 0.5 µm.
Reprinted with permission from Madden and Henson, 1997. From a paper on microwave
accelerated decalcification by Madden and Henson, Department of
Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC.
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