Intravital microscopy was performed in dorsal skin-fold window chambers installed on DPE-GFP or GREAT mice inoculated with MC38-H2B-mApple tumors. Mouse macrophages and/or vasculature were labeled with Pacific Blue ferumoxytol and dextran, respectively. AF647-aPD-1 (200 μg) was delivered i.v. and its tumor distribution was observed using an Olympus FluoView FV1000MPE confocal imaging system (Olympus America), as described previously (44 (link)). Pacific Blue, GFP/YFP, mApple, and AF647 were imaged sequentially using 405, 473, 559, and 635 nm lasers and BA430-455, BA490-540, BA575-620, BA575-675 emission filters with DM473, SDM560, and SDM 640 beam splitters, all sourced from Olympus America. Time lapse images were acquired continually over the first hour following AF647-aPD-1 injection, after which the mice were allowed to recover before subsequent imaging.
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Intravital Imaging of Tumor PD-1 Dynamics
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Corresponding Organization : Harvard University
Other organizations : Dana-Farber Cancer Institute
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Variable analysis
independent variables
- Mouse strain (DPE-GFP or GREAT mice)
- Tumor inoculation (MC38-H2B-mApple tumors)
- Labeling of macrophages and vasculature (Pacific Blue ferumoxytol and dextran)
- Delivery of AF647-aPD-1 (200 μg i.v.)
- Tumor distribution of AF647-aPD-1
- Intravital microscopy performed in dorsal skin-fold window chambers
- Imaging using Olympus FluoView FV1000MPE confocal imaging system
- Sequential imaging of Pacific Blue, GFP/YFP, mApple, and AF647 using specific laser lines and emission filters
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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