Alexa 488⦁rb∝FITC

Immuno: rb∝Neurotensin (1:8k) labeled with Alexa 488⦁rb∝FITC (1:2K, Invitrogen)—to be used in double labeled series, in combination with a streptavidin conjugate. We also have Cy 2⦁ms∝FITC (1:2k, Jackson), which works just as good as the Alexa, and Dylight 488⦁gt∝FITC (Rockland), that's being tested now.
Image: 20 panels at 10x, 1024x1024 (x20), 8-bit
Acquisition: 488 line at 1.8%, 1.5 Airy, Gain 569, Offset 0, Speed 6, Avg 4 (31 sec/panel; 10m 30 sec total)
Tiling: 20% overlap; Rotation Correction 0.8087º (calibrated from Zeiss-provided macro from within Zen)--& the Zeiss software still wouldn't stitch
Stitching: PS 5 photomerge, auto mode.
Image Processing: In PS Assigned profile Prophoto and converted to Adobe '98. Flattened, Cropped and resized. Colorized with Curves. Sharpened with Unsharp Mask. Reduced size, converted to sRGB and saved a copy as a JPEG

Conclusion: Great staining, amplifies as much or more than streptavidin. However, 10x 1.5 Airy clearly doesn't capture all the data (when viewed at actual size of the original which is 14 x 11" at 300 dpi) and, unlike the NOS, would probably benefit greatly from acquiring in Z...

Neurotensin (1:8k) labeled with Alexa 488⦁rb∝FITC (1:2K)—to be used in double labeled series, in combination with a streptavidin conjugate.