Ix71 fluorescence microscope

FTIR transmission spectra were recorded to evaluate the binding
of thiols and PBs to the GaAs surface, as well as the efficiency of
antibody grafting for different architectures. The measurements were
performed under vacuum using a Bruker Vertex 70v spectrometer, equipped
with a RockSolid interferometer and a wide-range Globar IR source
covering 6000 to 10 cm^–1. The spectra (1000 scans)
were collected with a liquid-nitrogen-cooled mercury cadmium telluride
IR detector at 4 cm^–1 spectral resolution and an
aperture of 1.5 mm. The spectrum of a freshly etched GaAs substrate
was used as reference and subtracted from the spectra of biofunctionalized
samples.
The presence of fluorescence labeled antibodies or
bacteria immobilized on the surface of the biochips was analyzed using
an Olympus IX71 fluorescence microscope equipped with GFP filters
(excitation at 473 nm and emission at 520 nm), FITC (excitation at
495 nm, emission at 519 nm), and a DP71 digital camera. Six to eight
images were collected per sample at different sites with a 20×
magnification using Q-capture software (QImaging Corporation, Surrey,
BC, Canada). The number of antibodies/bacteria present on the surface
was estimated for each sample after analysis of the fluorescent images
with ImageJ software.^{39 (link)}

NCI-H460 cells were fixed with 4% paraformaldehyde for 10 min, permeabilized with 0.1% Triton X-100 for 10 min and blocked with 1% FBS for 1 h at room temperature. For LC3 puncta and lysosomes staining, cells were incubated with antibodies against LC3 or LAMP1 antibody overnight at 4°C, and with anti-rabbit or anti-mouse IgG-FITC antibody, respectively, at 37°C for 1 h. Images were acquired using an Olympus IX71 fluorescence microscope at ×400 magnification (Olympus Corporation). Data were analyzed using Image J software (version 1.35; National Institutes of Health).