Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry
Article Title: Overexpression of Receptor Tyrosine Kinase EphB4 Triggers Tumor Growth and Hypoxia in A375 Melanoma Xenografts: Insights from Multitracer Small Animal Imaging Experiments ‡
Figure Lengend Snippet: Representative images of PET, radioluminography, in vivo fluorescence imaging, and ex vivo (fluorescence) microscopy of A375-pIRES/-EphB4 tumor-bearing NMRI nu/nu mice and tumor cryosections. After intravenous injection [ 18 F]FDG, [ 64 Cu]Cu-ETS, and [ 18 F]FMISO clearly accumulate in A375-pIRES and A375-EphB4 tumors and can be visualized by small animal PET ( A – C ) and subsequent radioluminography ( D – F ). ( A – C ) PET data are illustrated as maximum intensity projection (MIPs) of tracer-dependent frame times ([ 18 F]FDG, 30–60 min p.i.; [ 64 Cu]Cu-ETS, 0–30 min p.i.; [ 18 F]FMISO, 180–240 min p.i.). ( D – F ) Subsequent to PET imaging, A375-pIRES/-EphB4 tumors were resected and distribution of PET tracers was analyzed by radioluminography in tumor cryosections (18 sections each in 3 tumor depth). ( G ) In vivo fluorescence imaging of A375-pIRES/-EphB4 tumor-bearing NMRI nu/nu mice 24 h post i.v. injection of AngioSense (750/790 nm). ( H ) Ex vivo fluorescence microscopy of A375-pIRES/-EphB4 tumor cryosections following i.v. injection of 30 mg/kg Hoechst 33342 exactly 1 min before sacrifice of mice. ( I ) Tumor cryosections were stained with CD31 and LYVE-1 specific primary antibodies as well as Alexa Fluor 488- and Alexa 546-conjugated secondary antibodies to determine blood and lymph vessel markers CD31 (colored in green) and LYVE-1 (colored in orange), respectively. ( J ) Subsequent to fluorescence microscopy, tumor cryosections were stained with H E to get detailed data on tumor tissue, tumor stroma, and adjacent mouse tissue. Abbreviations: b, brain; bl, bladder; gbl, gall bladder; h, heart; in, intestine; li, liver; ln, lymph node; lu, lung; t, tumor; ki, kidney; v, vein.
Article Snippet: Primary antibodies for CD31 and LYVE 1 were visualized by Alexa Fluor 488-conjugated goat anti-rat IgG (Thermo Fisher Scientific, Langenselbold, Germany, ; 10 µg/mL) and Alexa 546-conjugated donkey anti rabbit IgG (Thermo Fisher Scientific, , 10 µg/mL, LYVE-1), respectively, for 1 h at room temperature and subsequent fluorescence microscopy using the microscope AxioImager.A1 (Zeiss, Jena, Germany) and software AxioVision (Zeiss).
Techniques: Positron Emission Tomography, In Vivo, Fluorescence, Imaging, Ex Vivo, Microscopy, Mouse Assay, Injection, Staining