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"I started using fluorescent media in 2011/12 in the latter part of my Fine Art Degree as I was looking into contrasting opposites with nature and fluorescent colour screams 'man-made' to me, as does the metallic leaf I also started experimenting with," writes McNaught.
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Samples labeled with paFPs can be imaged in any non-fluorescent media, whereas some synthetic dyes require the use of reducing agents in the imaging buffer to photoswitch properly [ 4, 9, 10].
Cells were grown in low-fluorescence media [ 60] for fluorescent microscopy experiments.
Specimens were mounted in fluorescent mounting media (DAKO) and analyzed by fluorescence microscopy (Axiovert 135, Zeiss).
The air-dried slides were mounted in fluorescent mounting media containing DAPI (Prolong Gold; Invitrogen-Molecular Probes) and imaged with a Nikon Histological Microscope.
Coverslips with stained cells were mounted on a glass slide treated with Dako Fluorescent Mounting Media with 0.1 μg/mL of membrane stain FM4-64.
Nuclei were counterstained with DAPI (Invitrogen) for 10 min and mounted in fluorescent mounting media (Dako).
Slides were mounted with Dako fluorescent mounting media and a glass coverslip and allowed to dry overnight.
Nuclei were visualized with either TO-PRO (Invitrogen), or 4',6-diamidino-2-phenylindole (DAPI), and sections were mounted using fluorescent mounting media.
Secondary antibodies, fluorescein-conjugated (Alexa Fluor 563, Alexa Fluor 488 - Invitrogen) were added for 45 min. Nuclei were counterstained with DAPI (Invitrogen) for 10 min and mounted in fluorescent mounting media (Dako).
Slides were mounted in fluorescent mounting media (DAKO).
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