Ethical approval for this study was obtained from the Institutional Animal Care and Use Committee of Hoshi University (approval no. P21-039). A total of 8 female BALB/c mice (weight, 18–20 g; age, 8 weeks; Sankyo Labo Service Corporation, Inc.) were housed at 24°C and 55% humidity under 12/12-h light/dark cycle (lights on at 8:00 a.m.) with food and water ad libitum. siRNA lipoplexes with 20 µg Cy5-siRNA were administered intravenously to mice via the lateral tail vein (n=1/siRNA lipoplex). At 1 h post-injection of siRNA lipoplexes, mice were sacrificed via cervical dislocation; death was confirmed by cessation of heartbeat. Tissue (lung, heart, liver, spleen, and kidney) was analyzed by Cy5 fluorescence imaging using NightOWL LB981 NC100 system (Berthold Technologies GmbH & Co. KG), as previously described (21 (link)). The images were analyzed using IndiGo2 software (version 2.0.1.0) provided with the in vivo imaging system (Berthold Technologies). Following fluorescence imaging, tissue samples were frozen on dry ice and sliced into 16 µm sections. The localization of Cy5-siRNA was examined using a fluorescent microscope (Eclipse TS100-F; Nikon Corporation) with optical filter Cy5 HQ (excitation, 620/60 nm; dichroic mirror, 660 nm; emission, 700/75 nm; Nikon Corporation).
In vivo imaging system
Manufactured by Berthold Technologies
Sourced in Germany
The In vivo imaging system is a laboratory equipment used for non-invasive visualization and quantification of biological processes within living organisms. It utilizes advanced imaging techniques to capture real-time data on various physiological and molecular activities.
Automatically generated - may contain errors
Lab products found in correlation
Nightowl lb981 nc100 system, by Berthold Technologies (1 mentions)
Indigo2, by Berthold Technologies (1 mentions)
Eclipse ts100 f, by Nikon (1 mentions)
Balb c mice, by Sankyo Labo Service (1 mentions)
Indigo software, by Berthold Technologies (1 mentions)
Nightowl lb 983, by Berthold Technologies (1 mentions)
Spectramax m5, by Molecular Devices (1 mentions)
Passive lysis buffer (plb), by Promega (1 mentions)
Luciferase assay system, by Promega (1 mentions)
D luciferin, by Promega (1 mentions)
8 protocols using in vivo imaging system
1
Biodistribution of Cy5-siRNA Lipoplexes
2
Biodistribution of Fluorescent Nanoparticles in Tumor-Bearing Mice
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BAF312 was replaced with the lipophilic tracer Dir in the NPs to detect the biodistribution of NPs in MDA-MB-231 tumor-bearing female BALB/c nude mice due to the strong fluorescence emission of Dir in the NIR region (λex/λem: 748/780 nm). 2 × 106 cells were subcutaneously injected into the upper right backs of 4 weeks old mice. Once the tumors volume reached 200 mm3, the mice were administered 100 μl of Dir@CaP-NPs or Dir@cRGD-CaP-NPs at a Dir equivalent dose of 0.5 μg/ml intravenously. After injection, the in vivo imaging system (Berthold Technologies, Germany) was used to obtain the whole-body fluorescence images at the time points of 2 h, 12 h, 24 h, and 96 h. The mice were sacrificed and dissected 96 h after the injection to examine the NP distribution in the major organs (spleen, heart, liver, lung, and kidney).
3
BALB/c Mouse Leukemia Model and MZ1 Treatment
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All our research involving animal experiments were approved and licensed by the Animal Care and Use Committee of the Children’s hospital of Soochow University (CAM-SU-AP#: JP-2018-1). SPF-grade BALB/c mice were purchased from Ling Chang Biotechnology Co., Ltd. (Shanghai, China). Five-week-old female mice were randomly grouped (5 mice per group) after tail vein injection of 3 × 105 luciferase-labeled P388-D1 cells. Bioluminescence imaging tests were performed using an in-vivo imaging system (Berthold, Germany) to obtain a successful leukemia model. The leukemic mice were then given a daily intraperitoneal injection of 12.5 mg/Kg MZ1 or the same dosages of vehicle (5% Kolliphor®HS15). The mice’s body weight was measured every three days.
4
Biodistribution of Hydrophobic Fluorescence Label
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Dir (40757ES25,Yeasen, Shanghai, China), a hydrophobic fluorescence label, was encapsulated into CaP‐NPs and cRGD‐CaP‐NPs. Free Dir, Dir‐NPs, and Dir‐NPs‐cRGD were delivered by intravenous injection, and their biodistribution in mice was studied using an in vivo imaging system (Berthold Technologies Gmbh & Co. KG, Bad Wildbad, Germany, LB983). At 96 h post‐injection, the mice were sacrificed, and the individual organs and tumors were harvested and scanned. The images were analyzed by indiGo software.
5
Bioluminescence Imaging for Bacterial Infection
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Following each 12 or 24 h of reperfusion since bacteria were injected, BLI was performed using an in vivo imaging system (Berthold Technologies, Germany) before and immediately after the laparotomy. Imaging signals were quantified as maximum photons per second per centimeter squared per steradian (p/s/cm2/sr) within the regions of interest.
6
Bioluminescent Tumor Imaging in Mice
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About 50–100 mg omenta tissues were dissected and lysed with 500 µL passive lysis buffer (Promega, Madison, Wisconsin, USA). After centrifugation at 12,000 rpm for 10 min at 4°C, the supernatant was collected. Luciferase activity was measured using the Luciferase Assay System (Promega, E1500) according to the manufacturer′s instructions in a luminometer (Molecular Devices, SpectraMax M5).
Luminescence imaging
A nude mouse (∼25 g) was first anesthetized using 100 µl of chloral hydrate (4%). Then, D-luciferin (125 mg/g, Promega) in PBS (pH 7.4) was intraperitoneally injected 10 min prior to bioluminescent imaging in order to provide substrate for the luciferase-expressing cancer cells. The peritoneal tumor engraftment was imaged by the in vivo imaging system (NightOWL LB983; Berthold Technologies, Germany). Fluorescent signals (expressed in counts/s) from the images were calculated and all images were analyzed using the IndiGo software provided with the in vivo imaging system (Berthold Technologies).
Luminescence imaging
A nude mouse (∼25 g) was first anesthetized using 100 µl of chloral hydrate (4%). Then, D-luciferin (125 mg/g, Promega) in PBS (pH 7.4) was intraperitoneally injected 10 min prior to bioluminescent imaging in order to provide substrate for the luciferase-expressing cancer cells. The peritoneal tumor engraftment was imaged by the in vivo imaging system (NightOWL LB983; Berthold Technologies, Germany). Fluorescent signals (expressed in counts/s) from the images were calculated and all images were analyzed using the IndiGo software provided with the in vivo imaging system (Berthold Technologies).
7
In Vivo Biodistribution of Nanomaterials
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To encapsulate Dir to form Dir@NPs, Dir@NPs‐FA, and Dir@NPs‐FA/AbCD47 nanomaterials were used. In advance, HUH7 tumor cells were inoculated over the root of the right thigh of mice at the number of 2×106 cells, and when the tumors grew to 200–300 mm3, the mice were intravenously administered 100 µL of Dir@NPs or Dir@NPs‐FA or Dir@NPs‐FA/AbCD47 at a 0.5 µg/mL Dir dose. Since the lipophilic tracer Dir had a strong fluorescence emission in the near‐infrared region (λex/λem: 748/780 nm), it could be used to detect the biodistribution of NPs in tumor‐bearing nude mice. After injection, whole‐body fluorescence images were obtained at 8, 24, 48, and 72 h time points using an in vivo imaging system (Berthold Technologies, Germany). Mice were sacrificed and dissected 72 h after injection, to examine the distribution of NP in the major organs (spleen, heart, liver, lung, and kidney).
8
Targeted Tumor Imaging and Drug Delivery
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Free Dir, Dir NPs, and Dir NPs‐RGD of 200 µL (Dir 10 µg mL−1 equiv) was injected into the mice via the tail vein when the tumor volume reached ≈200 mm3. Fluorescence images were analyzed with an in vivo imaging system (Berthold Technologies, Germany) at 2, 4, 8, 24, and 48 h. Then, the mice were sacrificed after 48 h, and the organs were collected to measure the fluorescence intensity. In addition, ATO, ATO NPs@Au, and ATO NPs@Au‐cRGD were injected into the mice via the tail vein when the tumor volume reached ≈200 mm3. The concentrations of ATO were 1 mg kg−1. After 24 h of the administration, mice were sacrificed, and the main organs were collected to measure the drug distribution by ICP‐MS.
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