SERS measurements were carried out using a micro-Raman system based on an Olympus BX41 microscope (Nanobase, Korea). The excitation source was a He−Ne laser operating at λ = 633 nm, and the laser spot was focused on the sample through a 20× objective lens. The SERS signals were recorded using a thermodynamically cooled electron-multiplying charge-coupled device (Andor, UK) mounted on a spectrometer with 1200 groove/mm grating. A portable Raman device was purchased from Metrohm (Switzerland). A diode laser operating at λ = 638 nm was used as the excitation source. The baseline correction of Raman spectra was performed using NanoSpectrum XperRF C series software (Nanobase, Korea). The spectral analysis was performed using Origin Pro V8 software (OriginLab Corporation, MA, USA). Transmission electron microscopy (TEM) images were obtained using a Tecnai G2 F30 S-Twin microscope (OR, USA) operated at 300 kV. Absorbance spectra were obtained using Cytation 5 (BioTek, VT, USA), and dynamic light scattering (DLS) data were acquired using a Nano-ZS90 (Malvern, UK). RT-qPCR assays were performed using the LightCycler 96 instrument and analysis software (Roche, Switzerland).
Electron multiplying charge coupled device
Manufactured by Oxford Instruments
Sourced in United Kingdom
The Electron-multiplying charge-coupled device (EMCCD) is a type of image sensor used in scientific and industrial applications. It employs an electron-multiplying register to amplify the signal from low-light images, enhancing the camera's sensitivity and enabling the detection of very faint signals.
Automatically generated - may contain errors
Lab products found in correlation
Bx41 microscope, by Olympus (2 mentions)
Originpro v8, by OriginLab (1 mentions)
Cytation 5, by Agilent Technologies (1 mentions)
Lightcycler 96 instrument, by Roche (1 mentions)
Ix 71 inverted microscope, by Oxford Instruments (1 mentions)
Eclipse ti, by Nikon (1 mentions)
Xe 100 microscope, by Park Systems (1 mentions)
4 protocols using electron multiplying charge coupled device
1
Plasmonic Raman Spectroscopy Characterization
2
Live-cell Imaging of GFP-labeled Neurons
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For live-cell imaging, culture medium was replaced with pre-warmed (37 °C) HEPES-buffered saline (HBS) (119 mM NaCl, 5 mM KCl, 2 mM CaCl2, 2 mM MgCl2, 30 mM glucose, 20 mM HEPES at pH 7.4). Wide-field fluorescence images were acquired using an Olympus IX-71 inverted microscope with UApo 150×1.45 NA oil immersion objective lens, an iXon electron-multiplying charge-coupled device, and an MS-2000 XYZ automated stage. During imaging, neurons were maintained at 37 °C and 60% humidity in an incubation chamber on the microscope. A 488 nm argon ion laser was used for the excitation of GFP. A 525/30 bandpass filter (Semrock) was used to detect the emission from GFP. Time-lapse images were taken in a single plane at 10 fps using streaming acquisition mode in MetaMorph software. For nocodazole experiments, the same dendrites were imaged first in HBS and after the treatment with 5 μg/ml nocodazole for 4 h.
3
Thermophoretic Aptamer-Assisted EV Assay
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Isolated EVs (5 μl) were mixed with 2 mM MgCl2, 7.5% PEG, and 100 nM aptamers targeting EpCAM or CD63 (Sangon Biotech, China) (table S6) in a final volume of 10 μl. The samples were immediately loaded in an 80-μm-thick microchamber (made by sandwiching an 80-μm-thick spacer between a bottom glass slide and a top sapphire slide) for pTAS detection without pretreatment. The microchamber was placed on an inverted fluorescent microscope (ECLIPSE Ti, Nikon, Japan), and the thermophoretic accumulation was achieved by focusing an infrared laser (1480 nm, 15 mW, Changchun Laser Optoelectronics Technology, China) on the glass slide for 10 min. The images before and after laser irradiation were captured by a 40× objective and an electron-multiplying charge-coupled device (Andor, UK).
4
SERS Characterization of Nano-Scaled Samples
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SERS spectra were acquired using a homemade micro-Raman system based on an Olympus BX41 microscope (Olympus, Japan). A He-Ne laser with 633 nm radiation (Melles Griot, Australia) was used as the excitation source and the laser light was focused on the sample through a 50× objective (Mitutoyo, Japan) of the microscope. The spectra were measured with a thermodynamically cooled electron-multiplying charge-coupled device (Andor, UK) mounted on the spectrometer with a 1200 groove/mm grating (Dongwoo Optron, Korea). A holographic notch filter was used to reject the laser light (Kaiser Optical Systems, USA). The acquisition time was 1 min. SEM images were obtained from a Nova230 microscope (FEI Company). AFM images were obtained from a Park Systems XE-100 microscope.
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