Cultured cancer cells were labeled with CM-DiI (Invitrogen, USA) before injection. Cultured cells were first collected and then washed three times with HBSS. Next, the cells were labeled with CM-DiI at 37 °C for 5 min, following by 15 min at 4 °C, and unincorporated dye was removed by rinsing three times with HBSS. The cells were then examined via fluorescence microscopy. Subsequently, 2-days-postfertilization (dpf) zebrafish larvae were mounted using 1.2% low-melting gel (Promega, USA), and then approximately 400 CM-DiI labeled cells were injected into the perivitelline space (PVS) of each larvae under a microinjector (Picosprizer III, USA). After injection, the xenografts were cultured at 34 °C. At 24 h postinjection (hpi), the zebrafish larvae with similar sizes of transplanted cells were collected for further analysis and then cultured at 34 °C until the end of experiments.
Picosprizer 3
Manufactured by Promega
Sourced in United States
The Picosprizer III is a precision pressure control system designed for the controlled application of small volumes of liquids, gases, or other materials. It provides precise, reproducible, and adjustable pneumatic control for microinjection, patch-clamp, and other microfluidic applications.
Automatically generated - may contain errors
4 protocols using picosprizer 3
1
Zebrafish Xenograft Tumor Modeling
2
Zebrafish Xenograft Transplantation and Imaging
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Adult zebrafish and embryos were cultured according to the breeding conditions. The zebrafish AB wild type was used in our study. Zebrafish handling procedures were approved by Nanjing Medical University. Cultured cancer cells were labeled with CM-DiI (Invitrogen, United States) before injection. The cells were then examined via fluorescence microscopy. Subsequently, 2-day-postfertilization (dpf) zebrafish larvae were mounted using a 1.2% low-melting gel (Promega, United States), and then approximately 400 CM-DiI-labeled cells were injected into the perivitelline space (PVS) of each larva under a microinjector (Picosprizer III, United States). After injection, the xenografts were cultured at 34°C. At 24 h post injection (hpi), zebrafish larvae with similar sizes of transplanted cells were collected for further analysis and then cultured at 34°C until the end of experiments. At 4 days post injection (dpi), the zebrafish larvae were also mounted using a 1.2% low-melting gel for the imaging experiments. Imaging experiments were performed via a stereomicroscope (MVX10, Olympus, Japan). The spatial resolution of the images was 1,600 × 1,200 (MVX10).
3
Zebrafish Xenograft Model for Cancer
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Cultured cancer cells were labeled with CM-DiI (Invitrogen, USA) before injection. Cultured cells were rst collected and then washed three times with HBSS. Next, the cells were labeled with CM-DiI at 37°C for 5 min, following by 15 min at 4°C, and unincorporated dye was removed by rinsing three times with HBSS. The cells were then examined via uorescence microscopy. Subsequently, 2-days-postfertilization (dpf) zebra sh larvae were mounted using 1.2% low-melting gel (Promega, USA), and then approximately 400 CM-DiI labeled cells were injected into the perivitelline space (PVS) of each larvae under a microinjector (Picosprizer III, USA). After injection, the xenografts were cultured at 34°C. At 24 hours postinjection (hpi), the zebra sh larvae with similar sizes of transplanted cells were collected for further analysis and then cultured at 34°C until the end of experiments.
In vivo imaging and quantitative analysis At 4 days postinjection (dpi), the zebra sh larvae were also mounted using 1.2% low-melting gel for the imaging experiments. Imaging experiments were performed via stereomicroscope (MVX10, Olympus, Japan) or confocal microscope using a 20X water-immersion objective (Fluoview 1000, Olympus, Japan). The spatial resolution of the images was 1600×1200 (MVX10) or 1024×1024 pixels (Fluoview 1000).
In vivo imaging and quantitative analysis At 4 days postinjection (dpi), the zebra sh larvae were also mounted using 1.2% low-melting gel for the imaging experiments. Imaging experiments were performed via stereomicroscope (MVX10, Olympus, Japan) or confocal microscope using a 20X water-immersion objective (Fluoview 1000, Olympus, Japan). The spatial resolution of the images was 1600×1200 (MVX10) or 1024×1024 pixels (Fluoview 1000).
4
Zebrafish Xenograft Imaging Protocol
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Cultured cancer cells were labeled with CM-DiI (Invitrogen, USA) before injection. Cultured cells were rst collected, then washed three times with HBSS. Next, the cells were labeled with CM-DiI at 37℃ for 5 min, following by 15 min at 4 ℃. Lastly, unincorporated dye was removed by rinsing three times with HBSS, then examined the cells by uorescence microscopy. 48-hpf zebra sh larvae were mounted by 1.2% lowmelting gel (Promega, USA), then about 400 CM-DiI labeled cells were injected into the PVS of each larvae under the micro-injector (Picosprizer III, USA). After injection, the xenografts were cultured at 34℃. At 24 h post injection (hpi), the zebra sh larvae with similar size of transplanted cells were picked up for the further analysis, then cultured at 34℃ until the end of experiments.
In Vivo Imaging And Quantitative Analysis At 4 days post injection (dpi), the zebra sh larvae were also mounted by 1.2% low-melting gel for the imaging experiments. Imaging experiments were performed by stereotype microscopy (MVX10, Olympus, Japan) or confocal microscope using 20X water-immersion objective (Fluoview 1000, Olympus, Japan).
The spatial resolution of the images was 1600 × 1200 (MVX10) or 1024 × 1024 pixels (Fluoview 1000).
In Vivo Imaging And Quantitative Analysis At 4 days post injection (dpi), the zebra sh larvae were also mounted by 1.2% low-melting gel for the imaging experiments. Imaging experiments were performed by stereotype microscopy (MVX10, Olympus, Japan) or confocal microscope using 20X water-immersion objective (Fluoview 1000, Olympus, Japan).
The spatial resolution of the images was 1600 × 1200 (MVX10) or 1024 × 1024 pixels (Fluoview 1000).
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