Nitric oxide content in root was quantified using DAF-FMDA under epifluorescence microscopy (Guo et al., 2003 (link); Chen et al., 2010 (link)). Five millimeter of root tip segments were soaked in 20 mM HEPES/NaOH buffer (pH 7.4) supplemented with 5 mM DAF-FMDA for 20 min. After washing three times with 20 mM HEPES/NaOH buffer, the root tips were analyzed microscopically (Nikon Eclipse 80i, Nikon, EX 460-500, DM 505, BA 510-560). The intensities of the green fluorescence from the root tips were quantified by measuring the average pixel intensity with Photoshop software (Adobe Systems) (Guo et al., 2003 (link)). Data are presented as the mean percentages of fluorescence intensity relative to that of the wild-type plants grown under the same conditions.
Ba510 560
Manufactured by Nikon
Sourced in Japan
The BA510-560 is a laboratory equipment product from Nikon. It is designed for use in research and scientific applications. The core function of the BA510-560 is to provide a stable and precise platform for microscopy and imaging tasks.
Automatically generated - may contain errors
Lab products found in correlation
Ex460 500, by Nikon (5 mentions)
Dm505, by Nikon (5 mentions)
Eclipse ti, by Nikon (3 mentions)
Neo scmos, by Oxford Instruments (3 mentions)
Ex540 25, by Nikon (2 mentions)
Plan apo 60 n a 1.4 objective, by Nikon (2 mentions)
Dm565, by Nikon (2 mentions)
Eclipse 80i, by Nikon (1 mentions)
Eclipse 80i upright microscope, by Nikon (1 mentions)
5 protocols using ba510 560
1
Nitric Oxide Quantification in Plant Roots
2
Epifluorescence Microscopy Imaging Protocol
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The samples were illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using an oil-coupled Plan Apo 60× N.A.1.4 objective (Nikon). UV cut-off filter blocks (TRITC: EX 540/25, DM565, BA605/55; GFP-B: EX460-500, DM505, BA510-560; Nikon) were used in the optical path of the microscope. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. Two ND filters (ND4, 25% transmittance for TRITC and ND1, 100% transmittance for GFP-B) were inserted into the illumination light path of the fluorescence microscope to reduce photobleaching of the samples.
3
Visualizing Tubulin Microtubule Assembly
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The droplet of tubulin solution after drying and stabilizing with taxol buffer, was illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using 2×, 20× objective lens (Nikon). UV cut-off filter block (GFP-B: EX460-500, DM505, BA510-560; Nikon) was used in the optical path of the microscope. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. ND filter (ND32, 3.1% transmittance for GFP-B) was inserted into the illumination light path of the fluorescence microscope to reduce photobleaching of the samples.
4
Azobenzene Microscopy Imaging Protocol
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The samples were illuminated with a 100 W mercury lamp and visualized by an epifluorescence microscope (Eclipse Ti, Nikon) using an oil-coupled Plan Apo 60× N.A.1.4 objective (Nikon). UV cut-off filter blocks (TRITC: EX 540/25, DM565, BA605/55; GFP-B: EX460-500, DM505, BA510-560; Nikon) were used in the optical path of the microscope. Images were captured using a cooled-CMOS camera (NEO sCMOS, Andor) connected to a PC. Two ND filters (ND4, 25% transmittance for TRITC and ND1, 100% transmittance for GFP-B) were inserted into the illumination light path of the fluorescence microscope to reduce photobleaching of the samples. In order to isomerize the azobenzene units, the flow cell was irradiated with the light passed through a UV-1A filter block (UV-1A: EX 365-410, DM400, BA400; Nikon).
5
Visualizing Nitric Oxide in Kas Roots
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The accumulation of NO in Kas roots was visualized using a NO probe, DAF-FM DA (4-amino-5-methylamino-2,7-difluorofluorescein diacetate). First, after washing with 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)-KOH (pH 7.4), the root apex was immersed in 200 μL DAF-FM DA (10 μM) for 30 min in dark. Excess fluorescence was removed by washing with HEPES-KOH (pH 7.4) three times, then visualized using an Eclipse 80i upright microscope with the following filters: EX 460-500, DM 505, and BA 510-560 (Nikon, Minato, Tokyo, Japan). The intensity of the fluorescence was calculated using Photoshop 7.0 (Adobe Systems Inc., San Jose, CA, USA).
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