Preparations were viewed in a Nikon epifluorescence microscope with FITC or TRITC
filter cubes, and with an Olympus Fluoview 1000 confocal microscope.
2+] in the organ of Corti, the explants were stained with TRITC
(red), DAPI (blue), and TUNEL (green).
The system will be used daily by graduate (PhD) students, postdoctoral fellows and laboratory technicians to collect images of cells, biomaterials and biological tissues that have been labeled with up to 4 different fluorochromes (typically YFP, Alexa 488, Alexa 555, Alexa 594, Alexa 647, GFP, cy3, TRITC
, Cy5, mKate, mTomato, mCherry, DAPI).
Preparations were examined using a Nikon epifluorescence microscope with FITC or TRITC
filter cubes, and photographed with a SPOT camera (Diagnostic Instruments, Inc.
Should come with DAPI, FITC, TRITC
and NIR fluorescence filters
Microscopy of the green fluorescence of Sytox Green and the red fluorescence of TRITC
, the secondary antibody for antiphospho-histone H3, SCP, TPep, and FMRFamide, was with a Biorad Radiance 2000 confocal microscope.
The slides were washed in PBS and treated for 45 min at 37 [degrees]C with FITC (fluorescein-isothiocyanate)-conjugated anti-rabbit IgG (immunoglobulin) antibody or TRITC
(tri-rhodamine-isothiocyanate)-con-jugated anti-goat IgG antibody (Boheringer Mannheim, Germany) diluted 1:50 in PBS, 5% Tween-20.
Subsequently, the specimens were rinsed and incubated for 12 h with two secondary antibodies mixed 1:1 (anti-mouse and anti-rabbit, conjugated with fluorophores CY5 and TRITC
Afterward, the slides were rinsed in PBS, incubated with anti-mouse antibodies conjugated to FITC (Sigma; 1:50 in PBS) for 3 h, washed in PBS, and finally incubated with a mixture of DAPI (4',6-diamidino-2-phenylindole, Sigma, 1:500 in PBS) and phalloidin conjugated to TRITC
(Sigma; 1:200 in PBS) for 30 min.
Samples were observed with an epifluorescent microscope equipped with dichroic filter blocks for FITC and TRITC
The eggs were observed for the absence of fluorescence within the perivitelline space (PVS) by using FITC or TRITC
excitation and barrier filters on a Nikon diaphot inverted microscope.