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1. |
Open
an OAAS package in clean area such as clean bench.
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2. |
Place
the appropriate amount of culture media containing
cells into each well of the OAAS plate. |
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3. |
Culture
cells according to the experimental protocol.
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4. |
Attached
cells and resorption area can be observed with a
phase contrast microscope, even during the process
of culture. |
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5. |
Remove
the culture media from the OAAS plate on completion
of culture. |
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6. |
Wash
the OAAS plate with saline and leave it in a solution
of 5% sodium hypochlorite for 5 minutes in order
to detach the cells. |
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7. |
Wash
the OAAS plate again with saline, and then dry it.
Observe the resorption area. |
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Image
Analysis for OAAS |
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You
can measure the degree of resorption by osteoclasts
in OAAS using one of the following methods. |
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Method
1: |
Get
images with a CCD camera attached to a microscope
and a capture or grabber card in your computer.
Then, measure the number and area of resorption
pits found in the images. The image analysis softwares used
frequently are Image-Pro Plus, Scion Image (for
MacOS and Windows), and Adobe Photoshop. |
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Method
2: |
Get images
with a digital camera attached to a microscope (low
magnitude, 1.25 × 10). Then, measure the number
and area of resorption in the images. |
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Method
3: |
Another
way to acquire images is to scan microscopic pictures
with a desktop scanner. |
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