These grid annotations were transferred to the single-channel TRITC
or FITC images, and images were exported that captured the content of each HPF.
Cocultures of hASMCs and CFBE (1 : 5 ratio) were stained with DAPI and observed either at the DAPI channel (a), FITC channel (b), or TRITC
Images were captured in black and white using appropriate filters for FITC, TRITC
and DAPI excitation, respectively.
For localization of RT in relation to ER, HeLa cells were treated with TRITC- conjugated antibodies to an ER marker calreticulin (e, f), and the overlay of FITC and TRITC
signals is shown.
A drop of the solution was placed on a slide preheated to 37[degrees]C and a sperm cell membrane integrity analysis was performed using a Nikon Eclipse 50i microscope equipped with a fluorescence analysis adapter (filter B-2EC, TRITC
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The cells were labeled with 0.1gg/ml Hoechst 33342, 100nM Mitotracker red and 1mg/ml wheat germ agglutinin conjugated to TRITC
(WGA--TRITC) to visualize nuclei, mitochondria, and plasma membrane respectively.
Secondary antibodies were used, TRITC
conjugated goat anti-rabbit IgG (1 : 200).
Yan, "Luminescence resonance energy transfer based on [beta]-[NaYF.sub.4]: [Y.sub.b], Er nanoparticles and TRITC
dye," Science in China.
Upon completion of this step, cells were washed in PBS (0.1 M, pH 7.4) for five minutes and incubated for 1 hour with anti-mouse and anti-rabbit secondary antibody produced in donkeys (Jackson, USA), conjugated to fluorophore AlexaFluor 488, AlexaFluor 594, FITC, and TRITC
, and kept under refrigeration with absence of light.
The ArrayScan captures fluorescent images in the 3 channels relevant to this assay (Hoechst, TRITC
for Sytox staining and FITC for NBD-PE staining) and performs automated image analysis to quantify cell nuclei, the number of dead cells, and NBDPE dye fluorescent intensity.
Samples were thawed at 37[degrees]C and then incubated with tetramethyl-rhodamine-isothiocyanate (TRITC
)-labeled monoclonal anti-vWF antibody directed to the D3 domain (Santa Cruz Biotechnology) at a final concentration of 20 nmol/L for 30 min at room temperature.