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).
Indigo2
Manufactured by Berthold Technologies
IndiGO2 is a software application developed by Berthold Technologies. It serves as a data acquisition and analysis tool for laboratory equipment.
Automatically generated - may contain errors
Lab products found in correlation
Nightowl 2 lb983, by Berthold Technologies (3 mentions)
Nightowl lb981 nc100 system, by Berthold Technologies (2 mentions)
D luciferin potassium salt, by Promega (2 mentions)
D luciferin, by Promega (2 mentions)
Eclipse ts100 f, by Nikon (1 mentions)
In vivo imaging system, by Berthold Technologies (1 mentions)
Balb c mice, by Sankyo Labo Service (1 mentions)
Chondroitin sulfate, by Fujifilm (1 mentions)
Nightowl 2 lb 983 system, by Berthold Technologies (1 mentions)
Nightowl lb983 system, by Berthold Technologies (1 mentions)
Nc 100 ccd deep cooled camera, by Berthold Technologies (1 mentions)
7 protocols using indigo2
1
Biodistribution of Cy5-siRNA Lipoplexes
2
Progesterone Primes Mice for PsV Challenge
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Mice were treated with 3 mg of progesterone subcutaneously four days before PsV challenge. The immunized mice were intravaginally pretreated with 50 µl of 4% nonoxynol-9 (N9, Igepal, Sigma) at six hours prior to PsV challenge, R4 and PBS injected female mice were used as controls. 20 µl of PsV preparation containing 2.5×105 to 1.0×107 IU of PsVs (encapsidated reporter plasmid pLucf) and 1% carboxymethyl cellulose (CMC, Sigma) was intravaginally instilled using a positive-displacement pipette. Forty-eight hours post-PsV challenge, mice were vaginally instilled with 0.4 mg of 5′-F-Luciferin (CellCyto Life Sciences). Three minutes later, luciferase signals were acquired for 5 min with a biofluorescence imaging (BFI) system of the LB 983 NightOWL II (Berthold Technologies), and analyzed with IndiGO 2 software (Berthold Technologies). Statistical significance was determined by one-tailed unpaired t-test.
3
In Vivo Imaging of Liver Metastasis
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For efficient delivery of siRNA into liver metastasis, lipoplexes with 50 µg Cy5.5-siRNA were administered intravenously into mice with HeLa-liver metastasis at 1 min after the intravenous injection of 1 mg chondroitin sulfate (Wako Pure Chemical Industries, Ltd.) at 10 days after inoculation of HeLa-Luc cells, as reported previously (sequential injection method) (15 (link),16 (link)). A total of 1 h after injection, the mice were sacrificed, and Cy5.5 fluorescent imaging of the tissues was performed using a NightOWL LB981 NC100 system (Berthold Technologies, Bad Wildbad, Germany). In Cy5.5 fluorescent imaging, the excitation and emission filters were set at 630/20 and 680/30 nm, respectively. The exposure time for fluorescence was 5 sec. A grayscale body-surface reference image was collected using a NightOWL LB981 CCD camera (Berthold Technologies). The images were analyzed using IndiGo2 software (version 2.0.1.0; Berthold Technologies) provided with the in vivo imaging system.
4
Bioluminescent Imaging of Mice
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After the mice were anesthetized with avertin, D-luciferin potassium salt was injected (200 mg/kg in PBS, i.p.; Promega). Ten to fifteen minutes after luciferin injection, intensity of the bioluminescence signal was measured for 1–60 s once a week using the Night OWL II LB983 system (Berthold Technologies). Imaging analyses were performed with the IndiGO2 software (Berthold Technologies). All values are shown as photons per second.
5
In Vivo Bioluminescence Imaging
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D‐luciferin potassium salt (Promega) was injected intraperitoneally. Ten to 15 minutes after injection, bioluminescence signals were measured using the NightOWL LB983 system (Berthold Technologies). Imaging analyses were undertaken with the IndiGO2 software (Berthold Technologies). All values are shown as photons per second.
6
Bioluminescence Imaging in Mice
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BLI was performed at the Vivoptic platform (Bordeaux University & CNRS, UMS 3767) using a NightOWL II LB 983 calibrated system equipped with an NC 100 CCD deep-cooled camera (Berthold Technologies™, Bad Wildbad, Germany). Mice were injected intraperitoneally with d -luciferin (2.9 mg in 100 µL PBS, Promega™, Madison, WI, USA,) and sedated 5 min later. Bioluminescence images (1 min exposure, 4 × 4 binning) and photographs (100 ms exposure) were taken 8 min after the luciferin injection. A low light emitting standard (Glowell, Lux Biotechnology Limited, Edinburgh, UK) was placed next to the animal during each image acquisition to provide a quality control. Pseudocolor images representing the spatial distribution of emitted photons were generated using IndiGO 2 software (Berthold Technologies™) and superposed to the corresponding photographs.
7
In vivo and ex vivo Bioluminescence Imaging
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In vivo bioluminescence imaging was performed as previously described [21 ]. For ex vivo bioluminescence imaging, resected lungs and livers were incubated with D‐luciferin (Promega, Madison, WI, USA) and the bioluminescence signal was measured with Night OWL II LB983 (Berthold Technologies, Bad Wildbad, Germany). The images were analyzed with the IndiGO2 software (Berthold Technologies). All values were shown as photons per second.
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In vivo bioluminescence imaging was performed as previously described [
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