Biology Reference
In-Depth Information
Differentiate the slides in the lithium carbonate solution for
●
30 s and in the 70% ethanol until there is a sharp contrast
between the blue of the white-matter and the colorless gray-
matter.
Counterstain in eosin for 1 min and cresyl violet for another
●
1 min.
Dehydrate through 95% and 100% alcohol, clear in xylene, and
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mount the slides by coverslip.
For rat,
Take pictures at magnifi cation of 1.25× to see the whole brain
3.4. Photography
●
slice in H&E staining coronal sections with a scale bar.
Transfer pictures to computer, and save them in .tif format.
●
For pig,
Take pictures in the sliced brain sections with a ruler as refer-
●
ence by a digital camera.
Scan the Luxol fast blue stained coronal sections with the scale
●
bar.
Transfer pictures to computer, and save pictures in .tif format.
●
3.5. Measuring
Launch software (Image J, NIH), and load images.
Calibrate the distance, using the scale as a reference.
●
●
Click the freehand selection button, drag the cursor to outline
●
the interested area on the image, and choose Measure under
Analyze menu.
Measure bilateral caudate and ventricle size three times and get
an average size (
1
).
Expressed as a percentage of the ipsilateral/contralateral of
3.5.1. Caudate Atrophy
in Rats After ICH (Fig.
1
)
●
●
ventricle dilation, caudate atrophy (
3
).
Measure bilateral hippocampus three times and get an average
size (
4
).
Expressed as a percentage of the ipsilateral/contralateral of
3.5.2. Hippocampus
Swelling and Atrophy
(Fig.
2
)
●
●
swelling or atrophy.
Measure the bilateral white matter and hemisphere and multi-
plying by the thickness (8 mm) of the sections.
Measure three times and get a mean value.
3.5.3. Pig White Matter and
Hemisphere Measurement
(Fig.
3
)
●
●
Express the measurement as a percentage of the ipsilateral/
●
contralateral.
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