Mice were anaesthetized with isofluorane and injected intraperitoneally with 10 μl per g of body weight d-luciferin (15 mg/ml; Perkin-Elmer). Bioluminescence images, each typically 3 min, large binning, f/stop 1, were then taken 10–20 min post-luciferin injection, using an IVIS 200 series imaging system (Perkin-Elmer).
Ivis imaging system 200 series
Manufactured by PerkinElmer
Sourced in United States
The IVIS Imaging System 200 Series is a preclinical in vivo imaging platform designed for bioluminescent and fluorescent imaging applications. The system utilizes a highly sensitive charge-coupled device (CCD) camera to capture real-time images of small animal models.
Automatically generated - may contain errors
Lab products found in correlation
D luciferin, by PerkinElmer (5 mentions)
D luciferin reagent, by GoldBio (2 mentions)
Living image software, by PerkinElmer (1 mentions)
Living image 3, by PerkinElmer (1 mentions)
D luciferin, by Promega (1 mentions)
Cell lysis buffer, by Promega (1 mentions)
Digital sight ds qimc camera, by Nikon (1 mentions)
Eclipse ti u, by Nikon (1 mentions)
11 protocols using ivis imaging system 200 series
1
In Vivo Bioluminescence Imaging
2
In Vivo Bioluminescence Imaging
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Mice were anaesthetized with isofluorane and injected i.p. with 10 µl/g d -luciferin (15 mg/ml; Perkin-Elmer). Sequential bioluminescence images, each typically 3 min, large binning, f/stop 1, were then taken 10–20 min post-luciferin injection, using an IVIS 200 series imaging system (Perkin-Elmer). Light output was quantified at peak signal intensity, typically 14 or 18 min post-luciferin injection, using Living Image software (Perkin-Elmer).
3
Astrocyte Activation Dynamics in Mice
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Astrocyte activation in 10-week-old male Gfap-luc mice was quantified before (baseline) and at 2 h, 6 h, 24 h, 48 h, 72 h, and 96 h after i.p. administration of either 0, 63, or 250 μg/kg TNF-α (n = 7 per group). Brain bioluminescence was detected as described previously [27 (link), 35 (link)]. Briefly, Gfap-luc mice were anesthetized by inhalation of 2% isoflurane in 1 L/min oxygen, shaved on the head, and injected with 126 mg/kg D-luciferin (Promega, product ID E1601) in the tail vein. Three minutes later the animals were scanned with a charge-coupled device camera (IVIS Imaging System 200 Series, PerkinElmer) mounted on a dark box. Photon emission from the whole brain was measured using Living Image 3.2 software (PerkinElmer) in a region of interest (ROI) that was kept constant across mice. Bioluminescence coming from the ears was considered to be basal Gfap activity and was excluded from the ROI. Imaging signals were measured in physical units of surface radiance (photons/s/cm2/steradian [sr]).
4
In Vitro and In Vivo Bioluminescence Imaging
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BLI of cells and animals was performed with an IVIS Imaging System 200 Series (PerkinElmer) and quantitated with Living Image® software by measuring photon flux (photons/s/cm2/steradian) in regions of interest drawn around appropriate signals. For in vitro BLI, cells were treated with cell lysis buffer (Promega) and placed in 96-well black imaging plates, and 100 μL D-Luciferin reagent (1.5 mg/mL) (Gold Biotechnology) were added to each well and mixed well, and 30 sec later BLI was performed and the signal was acquired for 1 min. For in vivo BLI, anesthetized mice were injected intraperitoneally with 75 mg/kg of D-Luciferin and images were acquired 2–5 min after injection. Acquisition times were 2 min initially and were reduced in accordance with signal intensity to avoid saturation.
5
In Vivo Bioluminescence Imaging of Mice
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BLI of animals was performed with an IVIS Imaging System 200 Series (PerkinElmer, Waltham, MA, USA) and quantitated with Living Image® software by measuring photon flux (photons/s/cm2/steradian) in regions of interest drawn around appropriate signals. For in vivo BLI, anesthetized mice (n = 5) were injected intraperitoneally with 75 mg/kg of D-Luciferin, and images were acquired 2 to 5 min after injection. Acquisition times were 2 min initially, and were reduced in accordance with signal intensity to avoid saturation.
6
Visualizing GFP-Expressing Cells in Bioreactor
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For direct imaging of the GFP-mMSCs attached to the 160 cm2 column at the end of culture, the column was removed and fluorescence microscopy (Nikon Eclipse Ti-u with a Nikon Digital Sight Ds-Qimc camera, Japan) was used to image GFP expressing cells.
To image the whole bioreactor using the IVIS Imaging System 200 series (Caliper, PerkinElmer, MA, USA), the cells were fixed with 4% paraformaldehyde and were stained with 10 μg/mL propidium iodide (PI, Invitrogen) for 10 minutes. The bioreactor was then washed with PBS and stored on ice prior to imaging. As negative controls, bioreactors containing no cells underwent the same fixing and staining process. The bioreactors were imaged with an excitation filter of 520 nm and an emission filter of 620 nm. The thresholds were adjusted to remove background auto florescence.
To image the whole bioreactor using the IVIS Imaging System 200 series (Caliper, PerkinElmer, MA, USA), the cells were fixed with 4% paraformaldehyde and were stained with 10 μg/mL propidium iodide (PI, Invitrogen) for 10 minutes. The bioreactor was then washed with PBS and stored on ice prior to imaging. As negative controls, bioreactors containing no cells underwent the same fixing and staining process. The bioreactors were imaged with an excitation filter of 520 nm and an emission filter of 620 nm. The thresholds were adjusted to remove background auto florescence.
7
Bioluminescence Imaging of Cells and Mice
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BLI of cells and animals was performed with an IVIS Imaging System 200 Series (PerkinElmer) and quantitated with Living Image® software by measuring photon flux (photons/s/cm2/steradian) in regions of interest drawn around appropriate signals. For in vitro BLI, 105 H441, CB-MSC or fusion progeny cells were resuspended in 50 μL PBS and placed in 96-well black imaging plates, and 50 μL D-Luciferin reagent (1.5 mg/mL) (Gold Biotechnology) were added to each well and mixed well, and 30 sec later BLI was performed and the signal was acquired for 1 min. For in vivo BLI, anesthetized mice were injected intraperitoneally with 75 mg/kg of D-Luciferin and images were acquired 2–5 min after injection. Acquisition times were 2 min initially and were reduced in accordance with signal intensity to avoid saturation.
8
Longitudinal Bioluminescence Imaging of Tumor Burden
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Mice were imaged from first week to eighth week post-inoculation to assess tumor burden. Mice were anesthetized using isoflurane and imaged after 10 min of D-luciferin (1.5 mg/mouse, Perkin Elmer, Waltham, MA, USA) I.P. injection using an IVIS Imaging System 200 Series (Perkin Elmer, Waltham, MA, USA). All the images were acquired by auto exposure. Using Living Image 4.7.4 software, region of interest (ROI) was generated to cover whole body, and the total flux (p/s) was obtained.
9
In Vivo Bioluminescence Imaging
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All the mice were weekly imaged from 1st~8/9th week of post-inoculation to access the tumor burden. For this, mice were anesthetized using 3% isoflurane and given D-luciferin (1.5 mg/mouse, Perkin Elmer, Waltham, MA, USA) via intraperitoneal injection. Mice were imaged in an IVIS Imaging System 200 Series (Perkin Elmer, Waltham, MA, USA). Images were obtained by auto exposure, and to acquire the total flux (p/s), a region of interest was created to cover the whole body using Living Image 4.7.4 software.
10
Monitoring Transdermal Cy5.5 Delivery
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Anesthetized mice
received microneedle patches containing encapsulated
Cy5.5 on the right upper site of dorsal skin for different amounts
of time. The Cy5.5 fluorescence signal was monitored using the IVIS
Imaging System 200 Series (PerkinElmer Inc.).
received microneedle patches containing encapsulated
Cy5.5 on the right upper site of dorsal skin for different amounts
of time. The Cy5.5 fluorescence signal was monitored using the IVIS
Imaging System 200 Series (PerkinElmer Inc.).
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