Immunological Response to Experimental Intracerebral Hemorrhage: Morphological Assessments - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 618

Table 6
(continued)

III. Negative control: infrared dye-labeled antibody only

Ipsilateral Ipsilateral Contralateral Contralateral

700 nm 800 nm 700 nm 800 nm

22192 37614 10318 32018

22058 37273 10781 32887

15903 32885 10432 32397

20943 34226 10154 31201

29297 37505 9878 30348

22079 35901 10313 31770

a r real
b c corrected BOLD values indicates average value for column

Note : Scanning time varies depending on the scan area; cropping
unwanted or empty background shortens the scan times. To scan
2 × 2 cm 2 area requires approximately 15-20 min.

2.9. Quantitative
Image Analysis

Equipment
Odyssey infrared imaging system application software, version 3.0;
Li-Cor Bioscience, Lincoln, Nebraska USA.
Protocol
The scanned image saved as “Analysis” can be open using Odyssey
software.
1. Choose the “Background method” and keep it consistent
through all slides you are going to analyze.
2. In each specifi c region of interest (ROI), draw a “Feature”
(white squares in Fig. 6a , b).
3. Using a copy-paste command, place as many of these ROIs as
desired. In our experiments, CD3, CD8, CD4 cell signal inten-
sities were quantitated ipsilaterally in area surrounding hema-
toma and contralaterally in the basal ganglia corresponding to
where the ICH injury was prevalent. It is important to keep
the feature areas consistent as the acquired number of pixels
affects the result.
4. Each feature receives an individual ID number which is refl ected
in the “Report”. These reports are text fi les that can be opened
in standard Microsoft Excel spreadsheets. There are numerous