Scattering-based DF microscopy was conducted using an inverted microscope (ECLIPSE Ti-U, NIKON, Japan). A Nikon Plan Fluor oil iris objective (100×) was used with an adjustable NA (0.5–1.3) and a Nikon DF condenser for DF imaging. We obtained DF scattering images by using an Andor EMCCD camera (iXon Ultra 897, UK). The Image J software was used to analyze the collected DF images. Furthermore, we took single particle spectra of AuNSs and HAuNSs by using an Andor spectrometer (SHAMROCK303i, SR-303I-A, UK) equipped with an Andor CCD camera (Newton DU920P-OE, UK). The scattered light from the AuNSs was collected by an objective lens and sent to the entrance of the spectrometer for taking a spectrum. We then dispersed the scattered light by a grating (300 l/mm) inside the spectrometer and detected by the Andor CCD camera (Newton DU920P-OE, UK). A background spectrum was obtained at an area without nanoparticles. Finally, we used Matlab programs specially designed for this study to conduct data analysis.
Plan fluor oil iris objective 100
Manufactured by Nikon
The Plan Fluor oil iris objective (100×) is a high-magnification microscope objective designed for use in various laboratory applications. It provides a 100× magnification and is optimized for use with oil immersion samples. The objective features a plan-apochromatic lens design, which helps to ensure sharp, high-contrast images across the field of view.
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3 protocols using plan fluor oil iris objective 100
1
Scattering-based Dark-Field Microscopy of Nanoparticles
2
Scattering-based Dark-field Microscopy of Nanoparticles
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We performed scattering-based dark-field (DF) microscopy using an inverted microscope (ECLIPSE Ti-U, NIKON, Japan). In the DF mode, we used a Nikon Plan Fluor oil iris objective (100×) with an adjustable numerical aperture (NA, 0.5–1.3) and a Nikon DF condenser for DF imaging. To obtain DF scattering images with high quality, we used an Andor EMCCD camera (iXon Ultra 897, UK). We analyzed the collected DF images with the Image J software. Furthermore, single particle spectra of AuNSs and AuNCs were taken by using an Andor spectrometer (SHAMROCK303i, SR-303I-A, UK) equipped with an Andor CCD camera (Newton DU920P-OE, UK). We collected the scattered light from AuNPs by an objective lens and sent to the entrance of the spectrometer for taking a spectrum. The scattered light was then dispersed by a grating (300 l/mm) inside the spectrometer, and detected by the Andor CCD camera (Newton DU920P-OE, UK). We obtained a background spectrum at an area without nanoparticles. Finally, Matlab programs specially designed for this study were used to perform data analysis and to obtain single particle spectra.
3
Dark-Field Spectroscopy of Plasmonic Nanoparticles
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We performed scattering-based DF microscopy using an inverted microscope (ECLIPSE Ti-U, NIKON, Japan). A Nikon Plan Fluor oil iris objective (100×) with an adjustable numerical aperture (NA, 0.5 -1.3) and a Nikon DF condenser were used for DF imaging. To obtain high-quality DF scattering images, we used an Andor EMCCD camera (iXon Ultra 897, UK). The collected DF scattering images were analyzed via ImageJ and Matlab. After obtaining the DF images, the DF scattering spectra of the individual gold nanoplates (AuNPs) were taken using an Andor spectrometer (SHAMROCK303i, SR-303I-A, UK) equipped with an Andor CCD camera (Newton DU920P-OE, UK). The light scattered from AuNPs was collected via an objective lens and directed to the spectrometer to obtain a spectrum. The scattered light was dispersed by a grating (300 lines/mm) inside the spectrometer and detected using the Andor CCD camera (Newton DU920P-OE, UK). Finally, a background spectrum was taken at an area without nanoparticles.
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