Fellow DHIGers:
I hope you are healthy and doing well.
The main reason for this note is to let you know about three multifocal-plane virtual slides of liver (
UM237-198, UCSF67-273, and UCSF169-271) which were recently added to the
PNWU Virtual Microscope database, but before describing the slides, I have a couple of requests.
First, although I've retired from my faculty position at Pacific Northwest University of Health Sciences, PNWU has granted me short-term access to my old microscope, and I'm 'keeping an eye out' for good slides to scan. If you have really good glass slides that you might loan me for scanning, could you please let me know? In addition to having the virtual slide available to everyone via the
PNWU Virtual Microscope, I could provide whoever lent me the glass slide with a copy of the resulting virtual slide. It takes several weeks to do a high-resolution multi-focal plane scan, so I'll only be able to scan a few slides and I'll have to be selective ... but I would like to get a few more slides scanned before I've worn-out my welcome at PNWU. I would be interested in any really great slide, but I would be especially interested in:
- brain stained by the Golgi method. Any cerebral cortex would be great, but ideally we are looking for a rodent brain (so we can match it to the Allen Institute atlas) sectioned to include cerebral cortex and cerebellum.
- liver perfused antemortem with Monastral blue B. The UCSF virtual slideset includes a single-focal-plane 40x version of such a slide (UCSF-282), but I'd like to try scanning the slide at higher resolution in multiple focal planes. Steve Rosen gave me one of the UCSF glass slides, but my slide has too little liver to be useful.
- thyroid gland stained specifically for parafollicular cells/C-cells (e.g., stained immunohistochemically for calcitonin).
- liver stained for bile canaliculi and lightly counterstained. Our recently-scanned slide of bile canaliculi (UM237-198, described below) is not ideal because it isn't counter-stained and orientation on the slide is difficult; as a result, I'm looking for a better slide to demonstrate bile canaliculi.
Second, I am worried about the persistence of the PNWU Virtual Microscope once I've worn-out my welcome at PNWU, and the chances of this tool surviving would be greatly enhanced if it were adopted by another institution. If you (
and your IT department) might be interested in hosting this tool, could you please let me know? The software is available for free under a GNU GPL3 license, and I would be happy to work with you to get the system up-and-running (
I'm now an "old retired guy"). I'm biased, but I believe that high-resolution multifocal-plane virtual slides are valuable teaching tools (
e.g., see the liver slides described below), and I think it would be a shame if this resource disappeared for want of a home. The
Viewer (
http://viewer.pnwu.edu) can be accessed directly (
presumably, this is what you would do in histology labs), but
SlideBox (
http://slidebox.pnwu.edu) is the entry-point for someone who is teaching histology. In addition to the multifocal-plane virtual slides, the Virtual Microscope also displays single-focal-plane slides, and most of the slides in the PNWU database originally came from the VMD and the University of Michigan databases (
in SlideBox, move your mouse over: "Limit slides by:" "Source").
The main reason for this note is to make you aware of three slides which were recently added to the PNWU Virtual Microscope's database:
UM237-198, UCSF67-273, and
UCSF169-271.
UM237-198 is a scan of a glass slide provided by Michael Hortsch (
from the University of Michigan glass-slide teaching set). It is stained for bile canaliculi and is 20µm thick (
41 focal-planes separated by 0.500µm), so focusing up-and-down can give a sense of the 3-dimensional organization of the bile canaliculi (
try clicking on "Cycle through focal planes" when at zoom-level 7, 8 or 9). Because there is no counter-stain, it is hard to get oriented and using this slide for teaching will require providing the students with more help than usual. I've written sample lab instructions (
attached and also available online; the blue text in the PDF's text & images are links to the slide) which may make it easier for students to get oriented and to use this slide productively. Because there are so many focal planes, if you open this slide directly from within
Viewer (
by clicking on "Choose a new slide" and entering slide number "3011") or from
SlideBox (
limit slides by "Focal plane" → "Multiple focal planes", then click "Get list of slides"), you may want to speed-up focus cycling (
Menu → Settings → Change focus settings..." → set "Focal-plane cycling timer" to between ~100 and ~150); the first focus cycle will be slow (
as the Viewer loads the focal planes), but focal-plane cycling should go much faster thereafter.
UCSF67-273 is a scan of a glass slide provided by Steve Rosen (
from the UCSF glass-slide teaching set). This slide, which is stained with a silver stain (
reticular fibers are black) and counter-stained with van Gieson stain (
collagen fibers are dark brown-purple), demonstrates how the reticular fibers (
in the space of Disse) wrap around the hepatic sinusoids (
click on "Cycle through focal planes"); the dark "granules" within the hepatocytes probably are lysosomes, which usually cluster at the cells' apical pole (
adjacent to the bile canaliculi, which are not apparent). At
lower magnification, this slide can be used to visualize the arrangement of the hepatic sinusoids relative to the "plates" of hepatocytes (
a central vein is at 145000x140600 & a portal canal is at 152400x141200), as well as other aspects of liver histology, such as
sinusoids draining into a longitudinally-sectioned central vein (
which drains into a sublobular vein), and a
branch of the portal vein draining into sinusoids.
UCSF169-271, which also is from a glass slide provided by Steve Rosen, is a Mallory-trichrome-stained section of liver. Hepatic lobules can be seen both in
cross-section (
central vein at: 158900x156840, and portal canals at: 159200x162300, 152900x157800, 156300x152700, 164000x152000, & 164000x161400), and in
longitudinal section (
two longitudinally-sectioned central veins at: 56300x138100 & 150000x129900 drain into a sublobular vein at lower-right, at 147600x139000, while the large conducting portal triad at upper-right, at 162000x131000, supplies the longitudinally-sectioned portal canal at 162000x140600 and a portal canal that is out of the plane-of-section but is indicated by the line of darkly-staining hepatocytes at 153000x134400). Other examples of central veins draining into sublobular veins can be found at
46800x93700 and at
123860x186460. The aniline blue in Mallory's trichrome stain colors both collagen fibers and reticular fibers, so although they aren't as obvious as in the silver-stained slide (
UCSF67-273, see above), reticular fibers (
stained blue) can be seen adjacent to the hepatic sinusoids in this Mallory-stained slide (
e.g., focus up and down at 21200x127700, or at 160160x173100). As an aside, as you know, reticular fibers in the spleen also form hoops around the splenic sinuses (
another example of discontinuous sinusoidal capillaries), and the reticular fibers surrounding the splenic sinuses can be seen by focusing up-and-down in the slide of Mallory-trichrome-stained spleen (
UCSFx-027). On
UCSF169-271, with careful looking, it is possible to identify
bile canaliculi (
e.g., at 45634x143843, start at focal-plane=4 and step up-and-down between focal planes ~0 to ~10).
In addition to the new liver slides, some of our other multifocal-plane slides might be useful to you. A listing of all of the multifocal-plane virtual slides in the PNWU database can be obtained by opening
SlideBox, moving your computer mouse over "Limit slides by: "Focal plane", selecting "multiple focal planes", and then clicking on the "Get list of slides" button. I like all of these slides (j
ust like a mother, I'm partial to all of my kids), but possibly the most noteworthy are:
- UCSF172-485 (Brain, pituitary, etc.): osteocyte processes in osseous canaliculi (e.g., 97900x93300; click on "Cycle through focal planes") and megakaryocyte nuclei (e.g., 103380x86930 & 102720x87480, click on "Cycle through focal planes").
- UCSF32-70 (Rat lower jaw): osteoclasts: (e.g., 97800x63500 & 41680x72420), osteoblasts (92500x72000), and early bell-stage tooth (86400x60000).
- UM145-29 (Jejunum): enteroendocrine (enterochromaffin?) cells and plasma cells (e.g., 252800x281700).
- UM145-59 (Neonatal forearm): longitudinally-sectioned muscle spindle (136000x68000 (in pronator quadratus)) and cross-sectioned muscle spindles (in many of the longitudinally-sectioned muscles, e.g., 184450x74500 (in flexor digitorum profundus)).
I apologize for the length of this note. I hope that it is useful.
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Jim Rhodes
email:
JRhodes98903@gmail.com phone: 509-225-9810
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