Szx 10 fluorescent microscope

Angiogenesis and tumor invasion were evaluated as described previously [12] (link), [13] (link). Briefly, after transplantation, the embryos were examined under an Olympus SZX-10 fluorescent microscope 2 days postinjection (dpi). All of the embryos were then mounted in 3% methylcellulose (Sigma, USA) so that they were oriented in the correct position for imaging. Both bright field and fluorescent images were captured with a QImaging digital camera controlled with Image-Pro Express software. Images were merged using an Adobe Photoshop CS2 (Adobe, USA) software program. The GFP labeled tumor angiogenesis and the relative emitted RFP fluorescence derived from adoptively transferred tumor cells were analyzed by ImageJ software (NIH, Bethesda, USA).

After fixation, larvae were washed three times with PBS. Prior to anti-angiogenic assessment, both control- and treated-larvae were briefly screened for any morphological defects using an Olympus SZX10 microscope. To assess anti-angiogenic activity, embryos were screened for inhibition of intersegmental vessel growth using the following critieria: (a) absence of intersegmental vessel and/or (b) incomplete sprouting of intersegmental vessel from dorsal aorta (DA) to dorsal longitudinal anastomic vessel (DLAV) [37] (link). For hyaloid vasculature analysis, zebrafish lenses were dissected and mounted on depression microscope slides. The hyaloid vessel plexus surrounding the lens were examined by flourescence microscopy using an Olympus SZX10 microscope. To determine anti-angiogenic activity, treated-larvae were screened for inhibition of hyaloid vessel growth under the following critieria: (a) reduction of primary branches emerging from optic nerve head and/or (b) altered branching pattern. The amount of intersegmental and hyaloid vasculature per treated larvae was quantified by direct observation using an Olympus SZX10 fluorescent microscope.