Fluorescence imaging was performed on an Olympus VS120 slide scanner, equipped with an X-Cite 120 LED light source and a Hamamatsu Flash 4.0 sCMOS camera or a spinning-disk confocal microscope (UltraVIEW; PerkinElmer), comprising an inverted stand (Axiovert 200 M; Carl Zeiss Microimaging, Jena, Germany) and a spinning-disk confocal head (CSU-X1; Yokogawa Corporation of America) with Velocity acquisition software (PerkinElmer) and dual cameras: a Hamamatsu C9100-23B EMCCD camera and a Hamamatsu Orca-R2 camera. Fluorescence channels were acquired sequentially. DAPI fluorescence was acquired with a 405 nm diode laser and a 445 (W60), 615 (W70) dual bandpass emission filter. Red (Alexa 546) fluorescence a 561 nm diode laser and a 445 (W60), 615 (W70) dual bandpass emission filter.
C9100 23b em ccd camera
Manufactured by Hamamatsu Photonics
Sourced in United States
The C9100-23B EM-CCD camera is a high-performance scientific imaging device designed for low-light applications. It features an electron-multiplying CCD (EM-CCD) sensor that amplifies the signal before readout, enhancing the signal-to-noise ratio. The camera is capable of capturing images with high sensitivity and fast frame rates, making it suitable for various scientific and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Perfect focus system, by Nikon (2 mentions)
Uplansapo na 1.4, by Olympus (2 mentions)
Ultraview vox sdc microscope, by PerkinElmer (2 mentions)
Axiovert 200m, by Zeiss (1 mentions)
Vs 120 slide scanner, by Hamamatsu Photonics (1 mentions)
Csu x1, by Yokogawa (1 mentions)
Orca r2 camera, by Hamamatsu Photonics (1 mentions)
Axiovert 200m fluorescence microscope, by Zeiss (1 mentions)
Em ccd 9100 50 camera, by Hamamatsu Photonics (1 mentions)
Cellview, by Greiner (1 mentions)
Csu x1 spinning disk head, by Nikon (1 mentions)
5 protocols using c9100 23b em ccd camera
1
Fluorescence Imaging of Cellular Structures
2
Visualizing Lipid Droplet Dynamics in Pollen Tubes
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LDs’ movement was observed using a Perkin Elmer Ultraview VOX Spinning Disc Confocal Microscope with a 100 × oil immersion lens equipped with a Hamamatsu C9100-23B EMCCD camera. LDs stained with Nile Red was excited by a 561 nm argon laser, and emission was collected between 580 and 700 nm. Time-lapse images were recorded in 2 s time intervals for 5 min. Nile Red has an emission peak at 635 nm, so there is no or very little emission in the green channel. The fluorescence signal of LDs in pollen tubes was recorded using the red emission filter at both t = 0 s and t = 6 s, and we assigned a pseudo-color (green) at t = 6 s so that the signals collected at 0s and 6s can be compared to each other.
3
Multi-Modal Live-Cell Imaging
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Spinning disk confocal (SDC) imaging was performed at a PerkinElmer UltraVIEW VoX SDC microscope (Perkin Elmer, Waltham, USA) using a 60x Apo TIRF (NA 1.49) oil immersion objective and Hamamatsu C9100-23B EM-CCD camera. Stacks were acquired with a z-spacing of 500 nm. Live-cell imaging was performed at 37°C, 5% CO2, 40% humidity using multiposition imaging with an automated stage and the Perfect Focus System (Nikon, Tokio, Japan) for automated focusing at each time point with a time-resolution of 6 s - 5 min/frame.
Total internal reflection fluorescence (TIRF) live cell imaging was performed at objective type TIRF setup (Visitron Systems, Puchheim, Germany) based on a Zeiss Axiovert 200M fluorescence microscope equipped with a 100 x Zeiss Alpha Plan Apochromat (NA 1.46) oil immersion objective and Hamamatsu EM-CCD 9100–50 camera. The TIRF angle was manually controlled. Live-cell imaging was performed at 37°C 5% CO2, 40% humidity with a time-resolution of 5 s/frame.
Stimulated emission depletion (STED) imaging was performed at a λ = 775 nm STED system (Abberior Instruments GmbH, Göttingen, Germany), using a 100 x Olympus UPlanSApo (NA 1.4) oil immersion objective with 590 and 640 nm excitation laser lines at room temperature. Nominal STED laser power was set to ~60% of the maximal power of 1200 mW with 10 µs pixel dwell time and 15 nm pixel size.
Total internal reflection fluorescence (TIRF) live cell imaging was performed at objective type TIRF setup (Visitron Systems, Puchheim, Germany) based on a Zeiss Axiovert 200M fluorescence microscope equipped with a 100 x Zeiss Alpha Plan Apochromat (NA 1.46) oil immersion objective and Hamamatsu EM-CCD 9100–50 camera. The TIRF angle was manually controlled. Live-cell imaging was performed at 37°C 5% CO2, 40% humidity with a time-resolution of 5 s/frame.
Stimulated emission depletion (STED) imaging was performed at a λ = 775 nm STED system (Abberior Instruments GmbH, Göttingen, Germany), using a 100 x Olympus UPlanSApo (NA 1.4) oil immersion objective with 590 and 640 nm excitation laser lines at room temperature. Nominal STED laser power was set to ~60% of the maximal power of 1200 mW with 10 µs pixel dwell time and 15 nm pixel size.
4
Live-cell confocal microscopy of vesicles
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Cells were seeded onto either 4-compartment (CELLview, Greiner BIO-ONE) or 1-compartment (MatTek Corporation) 35 mm-diameter glass-bottom imaging dishes. Prior to imaging, cells were washed twice and cultured in phenol red-free DMEMcplt. Live-cell time-lapse confocal microscopy was performed in a humidified incubation chamber at 37°C and 5% CO2 using a PerkinElmer UltraVIEW Vox Spinning-Disk microscope equipped with Yokogawa CSU-X1 spinning disk head, Nikon TiE microscope body, a Hamamatsu C9100-23B EM-CCD camera, an automated stage, and the Perfect Focus System (PFS). An Apo TIRF 60x/1.49 N.A. oil immersion objective was used. Multichannel images were acquired sequentially using solid state lasers with excitation at 445 nm for mTurquoise2, 488 nm for pHluorin, 514 nm for eYFP, 561 nm for CLIPATTO590, 640 nm for SNAPSIR647, and matching emission filters. For imaging of pHluorin-tagged CD63 expressing cells, the medium was supplemented with 25 mM Hepes (pH 7.4) to stabilize neutral pH. The imaging time interval of each experiment is specified in the figure legends.
5
Multimodal Imaging of Live Cells
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The majority of spinning disk confocal (SDC) imaging was performed at a PerkinElmer UltraVIEW VoX SDC microscope (PerkinElmer, Waltham, USA) using a 60× Apo TIRF (NA 1.49) oil immersion objective and Hamamatsu C9100-23B EM-CCD camera. Stacks were acquired with a z-spacing of 500 nm. Live-cell imaging was performed at 37°C, 5% CO 2 , 40% humidity using multiposition imaging with an automated stage and the Perfect Focus System (Nikon, Tokyo, Japan) for automated focusing at each time point with a time resolution of 3 min/frame. High time resolution SDC live-cell imaging (presented in Fig. 3) was performed at an SDC microscope based on Yokogawa CSU-W1 disc with Borealis illumination optimization and Nikon Ti2 microscope stand equipped with a 100× Apochromat TIRF (NA 1.49) oil immersion objective and two identical Andor iXon Ultra 888 Ultra EMCCD cameras. Live-cell imaging was performed at 37°C, 5% CO 2 , 40% humidity with a time resolution of 10 s/frame. STED imaging was performed at a λ = 775 nm STED system (Abberior Instruments GmbH, Göttingen, Germany), using a 100× Olympus UPlanSApo (NA 1.4) oil immersion objective with 590 and 640 nm excitation laser lines at room temperature. Nominal STED laser power was set to ~30% of the maximal power of 2,400 mW with 10 µs pixel dwell time and 15 nm pixel size.
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