Proliferation was gauged via in vitro bioluminescent imaging. We seeded HeLa-Luc cells in 96-well plates, and then imaged these cells at 48 h following ESRP1 or control vector transfection. Prior to imaging, wells were supplemented with 150 mg/mL (final concentration) D-Luciferin, and after 5 min a NightOWL LB 983 system (Berthold Technologies) was used for photon counting. Data were analyzed using IndiGO2 software.
Nightowl lb983 system
Manufactured by Berthold Technologies
Sourced in Germany
The NightOWL LB983 system is a versatile and advanced imaging platform designed for a wide range of applications in life science research. The system utilizes a high-sensitivity CCD camera and a state-of-the-art optical system to capture high-quality images and luminescence data from a variety of samples, including cell cultures, small animals, and other biological specimens. The NightOWL LB983 system is equipped with a temperature-controlled chamber and automated positioning capabilities, allowing for precise control over experimental conditions and sample handling.
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5 protocols using nightowl lb983 system
1
Bioluminescent Imaging of Cell Proliferation
2
Establishing Xenograft and Metastasis Models
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Male BALB/c athymic nude mice (4–6 weeks old) were purchased from the experimental animal center of Shanghai Institute for Biological Sciences. Mice were housed under standard conditions and maintained according to the institutional guidelines for animal care. To establish a subcutaneous xenograft model, 2 × 106 cells in 100 μl PBS were subcutaneously injected into the flank of mice. Tumors were removed after 4 weeks, and their size and weight were determined. The experimental metastasis model was established by injecting 4 × 106 cells into the tail vein of nude mice (10 per group). After 8 weeks, metastatic tumors were imaged with the NightOWL LB983 system (Berthold Technologies, Wildbad, Germany) following D -luciferin (Promega) injection. Animal procedures were authorized by the Institutional Animal Care and Use Committee of Harbin Medical University.
3
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.
4
Subcutaneous and Orthotopic Xenograft Models of Cholangiocarcinoma
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All experimental procedures involving animals were approved by The Animal Care and Use Committee of Harbin Medical University, China. Male BALB/c nude mice (aged 4–5 weeks) were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd. For the induction of subcutaneous xenograft tumors, the mice were randomly divided into the indicated groups (n = 6 per group). HCCC‐9810 cells (1 × 107) together with 1 × 106 macrophages were subcutaneously injected into the flanks of mice to form a subcutaneous model.
12 (link) All mice were well cared for with a free diet and comfortable padding. The tumor size was measured every 3 days with Vernier calipers, and the tumor volume was calculated using the following formula: volume = length × (width)2 × 0.5. The mice were sacrificed after 5 weeks, and the tumor volume and weight were assessed by double‐blinded evaluation. For the orthotopic xenograft model, 3 × 106 firefly luciferase‐transfected ICC cells and macrophages (1:1) were mixed and inoculated into the livers of nude mice. After 6 weeks, the tumor size was visualized with the NIGHTOWL LB983 system (Berthold Technologies).
12 (link) All mice were well cared for with a free diet and comfortable padding. The tumor size was measured every 3 days with Vernier calipers, and the tumor volume was calculated using the following formula: volume = length × (width)2 × 0.5. The mice were sacrificed after 5 weeks, and the tumor volume and weight were assessed by double‐blinded evaluation. For the orthotopic xenograft model, 3 × 106 firefly luciferase‐transfected ICC cells and macrophages (1:1) were mixed and inoculated into the livers of nude mice. After 6 weeks, the tumor size was visualized with the NIGHTOWL LB983 system (Berthold Technologies).
5
Orthotopic Xenograft and Metastasis Model
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Female BALB/c athymic nude mice (4–6 weeks old) were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd. Mice were housed under specific pathogen-free conditions and raised following institutional guidelines for animal care.
All experimental protocols involving animals were approved by the Animal Ethics Committee of Harbin Medical University, China.
The subcutaneous xenograft model was established by injecting 2 × 106 HCC cells in 200 µL PBS into the flanks of mice. Tumor volume was calculated at the 6th week when all mice were euthanized. Subcutaneous xenograft tumors were cut into 1 mm3 cubes and transplanted into the livers (left lobes) of mice to establish an orthotopic tumor model. Tumor size was assessed weekly with NIGHTOWL LB983 system (Berthold Technologies, Wildbad, Germany).
The pulmonary metastasis model was established as follows: 4 × 106 cells suspended in 0.15 mL PBS were injected into the tail vein of each mouse. After 6 weeks, the mice were euthanized, and their lungs were extracted.
All animals were euthanized to collect tumor tissues at the 6th week, and their livers were resected for IHC staining and ELISA.
All experimental protocols involving animals were approved by the Animal Ethics Committee of Harbin Medical University, China.
The subcutaneous xenograft model was established by injecting 2 × 106 HCC cells in 200 µL PBS into the flanks of mice. Tumor volume was calculated at the 6th week when all mice were euthanized. Subcutaneous xenograft tumors were cut into 1 mm3 cubes and transplanted into the livers (left lobes) of mice to establish an orthotopic tumor model. Tumor size was assessed weekly with NIGHTOWL LB983 system (Berthold Technologies, Wildbad, Germany).
The pulmonary metastasis model was established as follows: 4 × 106 cells suspended in 0.15 mL PBS were injected into the tail vein of each mouse. After 6 weeks, the mice were euthanized, and their lungs were extracted.
All animals were euthanized to collect tumor tissues at the 6th week, and their livers were resected for IHC staining and ELISA.
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