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Ff775 di01

Manufactured by IDEX Corporation

The FF775-Di01 is a lab equipment product manufactured by IDEX Corporation. It is designed for use in laboratory settings. The core function of this product is to provide a specific capability required for laboratory work, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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3 protocols using ff775 di01

1

Laser-Scanning Microscopy Protocol

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A half-waveplate and polarizing beam splitter set the excitation power sent to the microscope following pulse gating. The beam was expanded to fill the microscope objective back aperture, which was either a 25× objective (Olympus XLPLN25XSVMP2, 1.0 NA) for in vivo imaging or a 10× objective (Nikon CFIPlan10×, 0.25 NA) for cuvette experimentation. The excitation beam was scanned using a pair of galvanometer mirrors (Thorlabs, QS7XY-AG) conjugated to the objective back aperture using a scan lens (Thorlabs, SL50-2P2, f=50  mm ) and tube lens (Thorlabs, two AC508-400-C lenses in Plössl configuration, f=200  mm ) combination. The backscattered fluorescent signal was directed to a photomultiplier tube (PMT) (Hamamatsu, H10770PB-50) with a 775 nm cutoff dichroic filter (Semrock, FF775-Di01) and was further filtered with a 609/181 bandpass filter (Semrock, FF01-609/181-25), which immediately preceded the PMT photocathode. Imaging was controlled using a custom LabVIEW software. Although it may be necessary to synchronize EOM gating with the data acquisition sampling in some situations, this was not done for this work given our low sampling rate (160 kHz) relative to the gating frequency (1 MHz).
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2

Fourier-domain OCT and SLO Integration

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The imaging beam from the sample arm of the Fourier-domain (i.e., spectral domain) OCT system was optically integrated with the SLO subsystem via the second dichroic mirror (DM2, FF775-Di01, Semrock). We used a broadband light source with a 132-nm bandwidth centered at 860 nm (Broadlighter 860; Superlum Diodes Ltd., Cork, Ireland), which provides ~3.6 μm theoretical axial resolution in tissue. As Fourier-domain OCT detector, a custom spectrometer with a high-speed line CMOS camera (Sprint spL4096–140km; Basler, Ahrensburg, Germany) was used. The OCT system operates at speeds of 100,000 A-scan/s. Phase-variance OCT angiography (OCTA) was also obtained using this system (Zhang et al, 2015 (link)). Standard OCT and OCTA data processing have been applied to generate OCT and OCTA serial B-Scans.
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3

Fluorescence Imaging System Setup

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A half-waveplate and polarizing beam splitter set the excitation power sent to the microscope following pulse gating. The beam was expanded to fill the microscope objective back aperture, which was either a 25X objective (Olympus XLPLN25XSVMP2, 1.0NA) for in vivo imaging or a 10X objective (Nikon CFIPlan10X, 0.25NA) for cuvette experimentation. The excitation beam was scanned using a pair of galvanometer mirrors (Thorlabs, QS7XY-AG) conjugated the objective back aperture using a scan lens (Thorlabs, SL50-2P2) and Plössl tube lens (Thorlabs, AC508-400-C) combination. Backscattered fluorescent signal was directed to a photomultiplier tube (PMT, Hamamatsu, H10770PB-50) with a 775 nm cutoff dichroic (Semrock, FF775-Di01), and was further filtered with a 609/181 bandpass filter (Semrock, FF01-609/181-25) which immediately preceded the PMT photocathode. Imaging was controlled using a custom LabVIEW software. While it may be necessary to synchronize EOM gating with DAQ sampling in some situations, this was not done for this work given our low sampling rate (160 kHz) relative to the gating frequency (1 MHz).
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