Raman images were recorded by a confocal Raman microscope (alpha300 R; WITec, Ulm, Germany) equipped with a 488 nm excitation laser, provided by a single mode diode laser. Laser light was focused onto the sample with ca. 10 mW intensity by using a 60x water dipping objective (NA 1.0; Nikon, Tokyo, Japan). The samples were scanned in the x-y plane in a raster pattern at a constant stage speed. Spectra were collected with 1 μm step size and an integration time of 1 second. The back-scattered Raman signal was recorded by a spectrometer, equipped with a 600 lines/mm grating and a -67 °C cooled EMCCD camera (Newton 970; Andor, Belfast, UK) with a 1600x200 array of 16 μm pixels. All 3 batches were measured and a minimal of 10 cells per batch was imaged.
Newton 970
Manufactured by Oxford Instruments
Sourced in United Kingdom
The Newton 970 is a high-performance, back-illuminated scientific CMOS (sCMOS) camera designed for a wide range of scientific and industrial applications. It features a large 16.6 megapixel sensor, low noise, and high-speed data transfer capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Water immersion objective, by Olympus (2 mentions)
Wire 4, by Renishaw (2 mentions)
Alpha 300r, by WITec (1 mentions)
Alpha300ra, by WITec (1 mentions)
Shamrock 303i, by Oxford Instruments (1 mentions)
Raman microscope, by Renishaw (1 mentions)
Invia confocal raman microscope, by Renishaw (1 mentions)
Half wave plate, by Thorlabs (1 mentions)
Tecnai f20, by Thermo Fisher Scientific (1 mentions)
Su8010, by Thermo Fisher Scientific (1 mentions)
Spm 9700, by Shimadzu (1 mentions)
9 protocols using newton 970
1
Raman Imaging of Cellular Samples
2
Raman Imaging and Scanning Force Microscopy Analysis
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A combined Raman Imaging/Scanning Force Microscope System (WITec alpha 300 RA+, Germany) with WiTec Control FIVE 5.3 software was used for RAMAN measurements. Laser is equipped with a UHTS 300 spectrometer combined with a back-illuminated Andor Newton 970 electron multiplying charge-coupled device camera [resolution: ca. 300 to 400 nm (lateral) and 900 nm (z) with 100× objective].
The measurements were carried out at an excitation wavelength of λ = 532 nm and a laser power of 1 mW with 50 accumulations with an integration time of 0.5 s pixel−1. The samples were stacked on a glass slide. After adjusting the focus on the nonwovens at ×100 magnification, the Raman spectrum was recorded. A cosmic ray removal and a baseline correction were performed on all spectra. The peaks were then fitted with a Gaussian function using the built in routine of Origin 2016.
The measurements were carried out at an excitation wavelength of λ = 532 nm and a laser power of 1 mW with 50 accumulations with an integration time of 0.5 s pixel−1. The samples were stacked on a glass slide. After adjusting the focus on the nonwovens at ×100 magnification, the Raman spectrum was recorded. A cosmic ray removal and a baseline correction were performed on all spectra. The peaks were then fitted with a Gaussian function using the built in routine of Origin 2016.
3
Raman Microscopy for Molecular Analysis
4
Raman Spectroscopy for Flow Experiments
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Raman spectroscopy was performed
using a previously described home-built system42 (link) along with a commercial Raman microscope (InVia, Renishaw,
Inc.). Laser excitation for both was provided by a 632.8 nm HeNe laser.
Flow experiments were completed using the home-built system. The sample
was illuminated through a 40× water immersion objective (Olympus,
NA = 0.8), and the power measured at the sample was 1 mW. Raman scattering
was collected through the same objective and transmitted back to the
spectrograph and EMCCD (Newton 970, Andor). Spectra were recorded
in kinetic series with varying acquisition times. Maps of the SERS
electrodes were done with the commercial Raman microscope. A 20×
objective (Leica, NA = 0.4) was used to illuminate the sample, and
the laser power measured at the sample was ∼0.8 mW. A spectrum
was recorded at each point of the map with a 1 s acquisition time.
using a previously described home-built system42 (link) along with a commercial Raman microscope (InVia, Renishaw,
Inc.). Laser excitation for both was provided by a 632.8 nm HeNe laser.
Flow experiments were completed using the home-built system. The sample
was illuminated through a 40× water immersion objective (Olympus,
NA = 0.8), and the power measured at the sample was 1 mW. Raman scattering
was collected through the same objective and transmitted back to the
spectrograph and EMCCD (Newton 970, Andor). Spectra were recorded
in kinetic series with varying acquisition times. Maps of the SERS
electrodes were done with the commercial Raman microscope. A 20×
objective (Leica, NA = 0.4) was used to illuminate the sample, and
the laser power measured at the sample was ∼0.8 mW. A spectrum
was recorded at each point of the map with a 1 s acquisition time.
5
Raman Spectroscopy of Liquid Samples
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Raman spectroscopy was performed on the liquid samples placed into glass capillary tubes (sealed on both sides) using an inVia confocal Raman microscope (Renishaw, Gloucestershire, UK) equipped with a 20× long working distance objective (SLM Plan N, NA = 0.25, Olympus, Tokyo, Japan) and with a high sensitivity, ultra-low noise EMCCD (Electron Multiplying Charged Coupled Device) detection camera Newton 970 (Andor, Belfast, UK). The samples were excited with an argon laser (Stellar-REN, Modu-Laser, Centerville, UT, USA) operating at 457 nm (0.35 μW maximum laser power). Spectra were obtained at 1 cm−1 resolution (2400 lines/mm grating) in the spectral region of 325–1995 cm−1. The total acquisition time was about 50 s. Spectral acquisition and pre-processing were performed with the Renishaw WIRE 4.4 software from Renishaw.
6
Raman Imaging of Xanthophyll-Containing GUVs
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Raman spectroscopy was carried out using an inVia confocal Raman microscope (Renishaw, UK) with argon laser (Stellar-REN, Modu-Laser™, USA) operating at 457 nm (set at 70 μW power at the sample), equipped with 60x water immersed objective (Olympus Plan Apo NA = 1.2). Optical images of xanthophyll-containing GUV were obtained and elaborated with WiRE 4.1 software (Renishaw, UK). Based on such images, areas of approximately 10 μm × 10 μm for Raman scanning were selected and mapped with 0.5 μm spatial resolution. For the purpose of this study, all the images were recorded with a light intensity as low as possible. At each point of Raman image map the spectra were recorded with about 1 cm−1 spectral resolution (2400 lines/mm grating) in spectral region 350–1900 cm−1 using EMCCD detection camera Newton 970 from Andor, UK. Images were acquired with use of the Renishaw WiRE 4.1 system at high resolution mapping mode (HR maps). Acquisition time for a single spectrum was 0.1 s. All spectra were pre-processed by cosmic ray removing, noise filtering and baseline correction using WiRE 4.2 software from Renishaw, UK. At least 10 giant unilamellar vesicles with lutein and zeaxanthin were imaged and analyzed. Representative images are presented in the paper.
7
Raman Spectroscopy of Biological Samples
8
Characterization of WSe2 and WS2
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Raman and photoluminescence characterizations of WSe2 and WS2 were obtained by confocal Raman spectroscopy (LabRAM HR800) with a 532-nm laser wavelength. AFM (Shimadzu SPM-9700) was used to measure surface topology. The SHG experimental system mainly includes a femtosecond laser (Coherent Mirra 900F), electron multiplying charge-coupled device (Andor Newton 970), and spectrometer (Andor 500i). Scanning electron microscopy (SEM; SU8010) and transmission electron microscopy (TEM; Thermo Fisher Scientific Tecnai F20) were used to observe the structure of the devices.
9
Raman Spectroscopy of Biological Samples
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