A 7 mm diameter optical probe (EMVision, Loxahatchee, Florida) is used to deliver 80 mW of power from a 785 nm diode laser (Innovative Photonics Solutions, Monmouth Junction, New Jersey). Raman signal is acquired by detector fibres in the probe and delivered to a near-infrared-optimised spectrograph (LS785, Princeton Instruments, Princeton, New Jersey), and recorded by a deep depletion, thermo-electrically cooled CCD (Pixis 400BR, Princeton Instruments). The 3D scanner component of Marginbot consists of two motors and servomotors (Tower Pro, Shenzen City, China), which is controlled by a laptop. A customised program code written in LabVIEW (National Instruments, Austin, Texas) and MATLAB (Mathworks, Natick, Massachusetts) can (i) reconstruct a 3D diagram of the specimen margin based from captured images of it from all angles by a camera (Genius, Doral, Florida), (ii) control the movement of motors and servomotors, and (iii) receive feedback from these components via a signal relay port (Belkins, Playa Vista, California).
Ls785
Manufactured by Teledyne
Sourced in United States
The LS785 is a spectrometer designed for spectroscopic analysis. It measures the intensity of light over a specific range of wavelengths. The core function of the LS785 is to detect and analyze the spectral characteristics of various materials and substances.
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4 protocols using ls785
1
3D Imaging and Raman Spectroscopy for Specimen Margin Analysis
2
Portable Raman Spectroscopy for Skin Analysis
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A portable Raman spectroscopy instrument was built with an 830-nm diode laser (PI-EC-830-500-FC, Process Instruments, UT, USA), an imaging spectrograph (LS785, Princeton Instruments, NJ, USA), and a charge-coupled device (CCD; PIXIS1024BRX, Princeton Instruments, NJ, USA). The illumination collection geometry using oblique angle (off-axis) incidence of laser and noncontact vertical Raman collection was determined for the increased effective sampling volume of the targeted layer while reducing the collection of background signals.
A filtered laser beam of 250 mW was focused on the ear skin with an incidence angle of 60°, forming an elliptical beam of 1 mm × 2 mm. Light emission from the measurement spot of the skin surface was collected with a custom-made, round-to-linear optical fiber bundle consisting of 61 fibers (LEONI Fiber Optics Inc., VA, USA) to fully cover the CCD height. The fibers were custom-drawn (200 μm core diameter, Fiberguide Industries) to minimize fiber background signal. The diameter of the fiber bundle was 2 mm at the probe tip with a custom filter (Alluxa, CA, USA) attached to reject Rayleigh light. A mechanical shutter was installed inside of the spectrograph to minimize vertical pixel smearing. A full-frame image of the CCD for 285 s was consecutively acquired every 5 min with the help of Lightfield software (Princeton Instruments, NJ, USA).
A filtered laser beam of 250 mW was focused on the ear skin with an incidence angle of 60°, forming an elliptical beam of 1 mm × 2 mm. Light emission from the measurement spot of the skin surface was collected with a custom-made, round-to-linear optical fiber bundle consisting of 61 fibers (LEONI Fiber Optics Inc., VA, USA) to fully cover the CCD height. The fibers were custom-drawn (200 μm core diameter, Fiberguide Industries) to minimize fiber background signal. The diameter of the fiber bundle was 2 mm at the probe tip with a custom filter (Alluxa, CA, USA) attached to reject Rayleigh light. A mechanical shutter was installed inside of the spectrograph to minimize vertical pixel smearing. A full-frame image of the CCD for 285 s was consecutively acquired every 5 min with the help of Lightfield software (Princeton Instruments, NJ, USA).
3
Raman Spectroscopy for Material Analysis
4
Single-Cell Raman Tweezers Imaging System
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The Raman tweezers system used in this work was described previously [17, 24] . Briefly, a laser beam at 780 nm is introduced into an inverted microscope (Nikon, TiS) that contains an external phase contrast system and an immersion objective (Plan Apo 60×, NA1.4) to form a single-beam optical trap. A spore or growing cell in an aqueous medium can be trapped ~10 µm above the bottom quartz coverslip. Backward Raman scattering light from the trapped cell exceited by the same laser was collected and focused on the entrance slit of a spectrograph (Princeton Instruments, LS-785) equipped with a back-illuminated deep depletion charge coupled device (Princeton Instruments, PIXIS 400BR).
The live-cell imaging of hundreds of individual cells was performed with phase contrast microscopy incorporated in Raman tweezers [17] . Phase contrast images were captured with a digital CCD camera (1,392 x1,040 pixels) at a rate of 1 frame per 15 s for up to 10 hours. A homemade auto-focus system was developed to actively lock in the focus of the objective, in which a diode laser at 650 nm was introduced to detect the distance change between the objective and the surface of the sample coverslip. The feedback electronic signal was added to a piezo attached on the objective to lock its position. The measured long-term stability of the focus locking along the z direction was ~10 nm.
The live-cell imaging of hundreds of individual cells was performed with phase contrast microscopy incorporated in Raman tweezers [17] . Phase contrast images were captured with a digital CCD camera (1,392 x1,040 pixels) at a rate of 1 frame per 15 s for up to 10 hours. A homemade auto-focus system was developed to actively lock in the focus of the objective, in which a diode laser at 650 nm was introduced to detect the distance change between the objective and the surface of the sample coverslip. The feedback electronic signal was added to a piezo attached on the objective to lock its position. The measured long-term stability of the focus locking along the z direction was ~10 nm.
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