The presence of Ki67-positive tumor cells and CD31-positive vessels in rat stroma tissue was analyzed using a fluorescence laser scanning system (TCS-SP2 scanning system and DM IRB inverted microscope, Leica®, Solms, Germany).
Microscopy was performed using a fluorescence laser scanning microscope (LSM710; Carl Zeiss, Jena, Germany), equipped with Plan-Apochromat 40×/1.3NA and 63×/1.4NA objective lenses, and Zeiss ZEN (2010) software.
Images are captured using a fluorescence image scanning unit (FMBIO III) instrument (Hitachi, Yokohama, Japan) and the ratios of red (live cells) and green (dead cells) fluorescence were calculated.
Two-photon excited fluorescence images of the macaque cone mosaic were obtained using a fluorescence adaptive optics scanning laser ophthalmoscope, overcoming the challenges of a low numerical aperture, imperfect optics of the eye, high required light levels, and eye motion.
The fluorescence intensity was measured at excitation wavelength of 485 nm and emission wavelength of 528 nm using a fluorescence microplate reader (Dual Scanning SPECTRAmax, Molecular Devices Co., Sunnyvale, CA, USA).
All images were optimized by using a fluorescence microscope or a laser scanning confocal microscope (Nikon, http://www.nikonusa.com/).
Depending on the sample size, 10,000 500,000 cells were acquired by scanning using a Fluorescence Activated Cell sorter (flow cytometry) (BD-FACS-Calibur, Franklin Lakes, NJ), and analyzed using Flojo software as described previously 33.
After washing with PBS-T, the membrane was scanned using a fluorescence imager to analyse FLAG and biotin acceptor peptide signals.
The gels are scanned using a fluorescence scanner, which specifically measures the emission wavelength of each Cy-dye.
After incubation with fluorescently labeled secondary antibodies (goat-anti-mouse IRDye 800 CW or goat-anti-rabbit IRDye 680 LT; LI-COR Biosciences, Lincoln, NE, USA) diluted 1 : 10 000, membranes were scanned using a fluorescence scanner Odyssey (LI-COR Biosciences).
