Fluorescence microscopy measurements were performed on an Olympus IX71 inverted microscope with an UPlanSApo × 60/1.2 W water immersion objective. A Ti:sapphire oscillator (Chameleon, Coherent) centering at 800 nm served as the two-photon excitation source. A dichroic mirror 715DCSPXR (AHF) and a short pass filter E700SP were used to separate the excitation and emission and the dichroic mirror 590DCXR (AHF) to split the fluorescence onto the green and red avalanche photodiode detectors (SPCM-CD 2969, PerkinElmer), selected with band pass filters D525/20m and D680/30m (Chroma). Data were acquired with a router (PRT 400)-coupled TCSPC card (TimeHarp200, PicoQuant) and analysed with the SymPhoTime software version 5.3 (PicoQuant).
Spcm cd 2969
Manufactured by PerkinElmer
Sourced in United States
The SPCM-CD 2969 is a single photon counting module developed by PerkinElmer. It is designed for detecting single photon events with high efficiency and time resolution. The core function of this product is to convert low-level optical signals into electrical pulses that can be processed and analyzed by other equipment or systems.
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Lab products found in correlation
2 protocols using spcm cd 2969
1
Two-Photon Fluorescence Microscopy Protocol
2
Microgel Hydrodynamic Radius Determination
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Dynamic light scattering was used to determine the hydrodynamic radius of the microgels in bulk aqueous solution. Measurements were performed with a light scattering setup consisting of a HeNe laser (633 nm, 35 mW, JDS Uniphase Corporation USA), goniometer (ALV/CGS-8F, ALV GmbH Germany), digital hardware correlator (ALV-5000, ALV GmbH Germany), two avalanche photo diodes (SPCM-CD2969, Perkin Elmer Inc. USA), light scattering electronics (ALV/LSE-5003, ALV GmbH Germany), an index-match bath filled with toluene and an external programmable thermostat (Julabo F32, Julabo GmbH Germany).
Samples were measured at 20 • C and the scattering angle was varied from 30 • to 100 • (5 • increments). Data were collected in dual-cross mode with an acquisition time of 120 s per measurement. To avoid multiple scattering and to ensure free diffusion, all samples were highly diluted. Data were evaluated utilizing the second order cumulant fit method. There-fore, the first cumulant was plotted against the square of the scattering vector and fitted with linear regression. The slope of the fit corresponds to the diffusion coefficient from which the hydrodynamic radius was calculated using the Stokes-Einstein equation.
Samples were measured at 20 • C and the scattering angle was varied from 30 • to 100 • (5 • increments). Data were collected in dual-cross mode with an acquisition time of 120 s per measurement. To avoid multiple scattering and to ensure free diffusion, all samples were highly diluted. Data were evaluated utilizing the second order cumulant fit method. There-fore, the first cumulant was plotted against the square of the scattering vector and fitted with linear regression. The slope of the fit corresponds to the diffusion coefficient from which the hydrodynamic radius was calculated using the Stokes-Einstein equation.
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