Tumor hypoxia was determined using the Hypoxyprobe-1 Plus Kit (Hypoxyprobe) as previously descrived13 (link),14 (link). Mice were injected intravenously with pimonidazole HCl (60 mg kg−1). Tumors were excised, and frozen sections were fixed with chilled acetone and stained with anti-pimonidazole adduct FITC-conjugated antibody according to the manufacturer’s instructions. Sections were imaged using an Axio Imager microscope (Zeiss) equipped with an AxioCam 506 monochrome CCD camera (Zeiss). Whole tumors were analyzed by light sheet fluorescence imaging using the Lightsheet Z.1 LSFM system (Zeiss).
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Quantifying Tumor Hypoxia via Hypoxyprobe
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Hypoxyprobe-1 Plus Kit
Axio Imager microscope
AxioCam 506 monochrome CCD camera
Lightsheet Z.1 LSFM system
Acetone
Anti antibody
Fitc
Frozen sections
Hypoxyprobe 1
Light
Mice
Microscope
Pimonidazole
Tumor hypoxia
Tumors
Corresponding Organization :
Other organizations : University of Pennsylvania
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Variable analysis
independent variables
- Pimonidazole HCl injection (60 mg kg^-1)
- Tumor hypoxia
- Fluorescence imaging of pimonidazole adducts
- Tumor excision
- Frozen sections
- Acetone fixation
- Staining with anti-pimonidazole adduct FITC-conjugated antibody
- Imaging using Axio Imager microscope and AxioCam 506 monochrome CCD camera
- Whole tumor analysis by light sheet fluorescence imaging using Lightsheet Z.1 LSFM system
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