Ivis spectrum ct

Mice were anesthetized with isoflurane (Patterson Companies, Inc, St. Paul, MN, USA), and 150 mg Luciferin/kg body weight were injected in the intraperitoneal cavity. Animal heads were shaved prior to imaging to avoid significant scattering/blocking of the optical signal. Animals were placed on the warmed In vivo Imaging System (IVIS® Spectrum CT, Perkin Elmer, Waltham, MA, USA) stage under 1.5% isoflurane and luminescence acquisition was taken 15 minutes after injection. Epi-fluorescence images using DsRed (570/620 nm) were also obtained. After IVIS®, the mouse was transferred to the MPI bed under anesthesia, and images were acquired using high sensitivity mode (3 T/m) in a FOV of 6 x 12 cm. Fiducials of known iron mass were within the same FOV in the MPI. The MPI bed was adapted to fit the CT stage to perform dual 3D imaging and anatomical co-registrations. Anatomic CT (IVIS® Spectrum CT, Perkin Elmer, Waltham, MA, USA) reference images were acquired on anesthetized animals (20 ms exposure time, 440 AI X-Ray filter). After in vivo imaging, mice were euthanized with isoflurane overdose and the liver, lungs, spleen, kidneys, and brain were collected and imaged via MPI and IVIS® epifluorescence.

For phantom fluorescence imaging,
phantoms were placed inside a preclinical IVIS (IVIS SpectrumCT, PerkinElmer)
and fluorescence intensity was recorded using the bandpass emission
wavelength of 680 nm and excitation wavelength of at 640 nm. The fluorescence
intensity, scale bars, and color maps were adjusted to the same levels.
For whole body fluorescence imaging, mice were imaged using the
same IVIS instrument (PerkinElmer IVIS Spectrum CT) at day 2 post-CLIO.
Prior to imaging, mice were shaved and depilated to remove all hair
that otherwise absorbs light and interferes with optical imaging.
Mice were anesthetized (Isoflurane 1.7%, O₂ 2 L/min). Mice
were imaged to detect the specific fluorescence of CLIO-AF647. The
fluorescence intensity, scale bars and color maps were adjusted to
the same levels. For ex vivo quantification of fluorescence intensity,
the femur, muscle, liver, spleen, and kidney were dissected and imaged
to detect specific fluorescence of CLIO-AF647. Fluorescence IVIS imaging
data were analyzed with Live Image 4.5.4 software.

Inflammation levels in the brain of Albino B6 mice, following i.c challenge, were detected by using in vivo Imaging System (IVIS SpectrumCT, PerkinElmer) and a fluorescent in vivo imaging agent (ProSense 750 FAST, NEV11171, PerkinElmer). At the day of imaging, the ProSense 750 FAST was reconstituted in PBS and each mouse was intravenously injected with 300 μl containing 4 nmol of the probe. 5–6 h post administration of the fluorescent probe, mice were transferred to the IVIS SpectrumCT (PerkinElmer) and scanned for fluorescence. During the scan, mice were kept under isoflurane anesthesia. Data obtained by IVIS analysis were subsequently analyzed with the “living image” software (PerkinElmer). The measured fluorescence was expressed as average radiant efficiency (p/s/cm²/sr)/(μW/cm²). Fluorescence measured on the back of each mouse served as background fluorescence and was subtracted by the fluorescence measured on the brain area.