DRS measurements were obtained between 400 and 1600 nm. Details on the measurement setup were reported previously.28 (link),29 (link) In short, the light of a halogen light source was transferred to the tissue by means of the illuminating fiber that is integrated into a fiber-optic probe. Two other integrated fibers were attached to two spectrometers, one covering the visual wavelength range (Andor Technology, DU420A-BRDD, 400 to 1000 nm) and one covering the near-infrared wavelength range (Andor Technology, DU492A-1.7, 900 to 1600 nm), both of which collected the photons after interacting with the tissue. The setup was controlled with LabVIEW® (Austin, Texas) software that performed a calibration from photon counts to diffuse reflectance and combined the output of both spectrographs to form one continuous spectrum from 400 to 1600 nm. For the calibration, a cap with Spectralon® (SRT-99-100, Labsphere, Inc., Northern Sutton, New Hampshire) at the bottom was used. The fiber distance between the emitting and collecting fiber was 1 mm.
Du492a 1
Manufactured by Oxford Instruments
The DU492A-1.7 is a laboratory equipment product manufactured by Oxford Instruments. It is a scientific instrument designed for specific technical functions. No further details about its core function or intended use are provided, as an unbiased and factual description cannot be given concisely without potentially making interpretations or extrapolations.
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5 protocols using du492a 1
1
Broadband Diffuse Reflectance Spectroscopy
2
Fiber-Optic Diffuse Reflectance Spectroscopy
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Fiber-optic point measurements were acquired with a measurement setup [shown in Fig. 2(b) ] including a broadband light source, a spectrometer for the visual wavelength region (Andor Technology DU420A-BRDD), and a spectrometer for the near-infrared region (Andor Technology, DU492A-1.7). A fiber-optic probe was attached to the measurement setup to measure diffuse reflectance spectra between 400 and 1600 nm [Fig. 2(d) ].18 (link),19 (link) The distance between the illuminating and collecting fiber at the tip of the probe was 1 mm. Before measuring, the setup was calibrated by acquiring a white reference measurement of Spectralon® (SRT-99-100, Labsphere, Inc., Northern Sutton, New Hampshire) in a calibration cap.
To acquire a DRS measurement, the probe was brought in contact with the tissue. During the probe measurements, a custom-made grid with holes [consisting of two pieces and shown inFig. 2(c) ] was placed on top of the tissue to gently fixate the tissue in the cassette and allow correlation between the measurement locations and H&E section afterward. The grid was 3-D printed and transparent. In addition, the grid stabilized the probe while ensuring contact between the probe and the tissue and restraining it from moving during measurements.
To acquire a DRS measurement, the probe was brought in contact with the tissue. During the probe measurements, a custom-made grid with holes [consisting of two pieces and shown in
3
Multimodal Imaging for Tissue Assessment
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The fiberoptic DRS system consisted of a Tungsten halogen broadband light source and two spectrometers; one for the visible domain (400–1100 nm, Andor Technology, DU420ABRDD) and one for the infrared domain (900–1700 nm, Andor Technology, DU492A-1.7)48 (link). A probe with two source-detector fiber distances of 2 and 6 mm was used to retrieve information from both superficial and deeper sampling depths. In DRS, the sampling depth (penetration depth of the detected photons) depends on the distance between the source and detector fiber. Even though the sampling depth is also influenced by the sample optical properties, it has been shown that the sampling depth is approximately equal to the source-detector distance49 (link). Ultrasound images were acquired using the portable Philips CX50 machine (Philips Research, Eindhoven, The Netherlands) in combination with the Philips L15-7io transducer (Philips Research, Eindhoven, The Netherlands), a high-frequency 15–7 MHz ultrasound transducer specially designed for superficial imaging.
4
Diffuse Reflectance Spectroscopy of Tissue
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Measurements were obtained with a system consisting of a broadband tungsten halogen light for illumination and two different spectrometers to record the diffuse reflectance spectra from the tissue. The first spectrometer contained a silicon detector resolving the visual light between 400 and 1100 nm (Andor Technology, DU420A-BRDD), and the second spectrometer contained an InGaAs detector resolving light in the NIR region from 800 to 1700 nm (Andor Technology, DU492A-1.7). 14 (link) The optical fibers guiding the light from the light source toward the tissue and the light reflected from the tissue toward the spectrometers are in contact with the tissue via a handheld probe. Multiple diffuse reflectance spectra were obtained for different distances between source fiber and detector fiber: 0.3, 0.7, 1, and 2 mm (Fig. 1). Different source-detector distances were used to obtain different sampling depths.
5
Diffuse Reflectance Spectroscopy System
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The DRS system consists of a Tungsten halogen broadband light source (360 to 2500 nm) with an embedded shutter and two spectrometers. Together, the two spectrometers cover the visible and near-infrared light range; the first spectrometer resolves Journal of Biomedical Optics 106014-2 October 2017 • Vol. 22 (10) light in the visible range 400 to 1100 nm (Andor Technology, DU420ABRDD), whereas the second resolves light in the near-infrared range 900 to 1700 nm (Andor Technology, DU492A-1.7) (Fig. 2). The spectrometers are controlled by custom LabView software (National Instruments) to acquire the data. The DRS system used in this study has been extensively described previously together with the calibration used for the system. 19, (link)20 (link) In all patients, the same probe, shown in Fig. 2, was used to obtain the DRS measurements. The probe consists of three optical fibers with a core diameter of 200 μm. One of the three fibers was used to transport the light from the source to the tissue, whereas the other two were used to transport the light from the tissue to one of both spectrometers. The distance between the two collecting fibers and the delivering fiber is 2 mm. This gives a mean sampling depth of about 2 mm (max 4 mm).
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