Biomedical Engineering Reference
In-Depth Information
2.5 In Vivo Imaging of the Oxidation Potential
After anesthesia 30
g of hydrocyanine solution was injected subcutaneously at the
site of implantation. After 30mins, fluorescent imaging was done in the near infrared
spectrum using in vivo imaging system (IVIS200, Xenogen, USA). The excitation
wavelength of hydrocyanines was 750nm and the emission wavelength was 840nm.
Acquired images were corrected for the background using image math tool of living
image software (version 4.3.1, Caliper life Sciences, 2012. In order to correct for the
background, a region of interest was selected from a mouse without implants after
addition of the fluorophore. The background value was calculated and automatically
subtracted from the images by the software.
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2.6 Monitoring Inflammatory Protease Activity
Protease activity dependent fluorescent sensor, Prosense 680 (PerkinElmer,
Germany) was used according to the manufacturer's recommendations. A dose of
2nm was injected intravenously into the mouse tail. Fluorescent imaging of the tis-
sue with implants was done after a period of 30min with an excitation and emission
wavelength of 680 and 700nm, respectively, using in vivo imaging system (IVIS
200). Similarly, the background of fluorescent images was subtracted using image
math tool of living image software as described above.
2.7 Visualization of Cell Growth Signaling
Lipase activity in the mouse tissue was monitored using ATX Red (L2010, Echelon
Biosciences Salt Lake City, UT). 10
g of ATX Red fluorophore was injected intra-
venously into the mouse tail. Whole animal imaging was done in the near infrared
spectrum at an excitation and emission wavelength of 775 and 800nm, respectively,
30mins after injection of ATX Red. Images were processed in the same way as
explained above.
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2.8 Imaging Interferon-
β
Induction in Transgenic Reporter Mice
Transgenic female BALB/c mice with a luciferase gene replacing the IFN-
coding
sequence on one allele were used for non-invasive in vivo imaging of the interferon-
β
β
induction [
32
]. 150
l of (30mg/ml) luciferin (PerkinElmer) was injected intraperi-
toneally. After 15min, luminescent imaging was performed using an in vivo imaging
system (IVIS 200).
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