Uhts 300

Manufactured by WITec

Sourced in Germany

The UHTS 300 is a high-performance ultra-high-throughput spectrometer designed for advanced spectroscopic analysis. It features a high-resolution optical system and a state-of-the-art detector, enabling rapid and precise data acquisition across a wide spectral range.

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Lab products found in correlation

Alpha300ra, by WITec (11 mentions) W plan apochromat, by Zeiss (7 mentions) Alpha 300r, by WITec (7 mentions) Control four, by WITec (6 mentions) Du401a bv, by WITec (4 mentions) Opus 7, by Bruker (4 mentions) Xtra 2, by Toptica (3 mentions) Oil immersion objective, by Zeiss (3 mentions) Du401a, by Oxford Instruments (3 mentions) Alpha 3000, by WITec (2 mentions) Du401 dd, by Oxford Instruments (2 mentions) Project v4, by WITec (2 mentions) Du401a br dd 352, by Oxford Instruments (2 mentions) Project four, by WITec (2 mentions) Newton du970n bv 353, by Oxford Instruments (2 mentions) Objective lens, by Nikon (2 mentions) Ec epiplan, by Zeiss (2 mentions) Charge coupled device camera, by Oxford Instruments (1 mentions) Matlab 2016b, by MathWorks (1 mentions) Igor pro 7, by Wavemetrics (1 mentions)

Spelling variants of this product

UHTS 300 spectrometer (9 citations) UHTS 300 Raman spectrometer (4 citations)

39 protocols using uhts 300

Confocal Raman Microspectroscopy Protocol

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We applied a confocal Raman microspectroscopy system (Alpha 3000, Witec, GmbH.). The microscope stage was equipped with a piezoelectric stage (UHTS 300, Witec, GmbH.). A green laser (λ_ex = 532 nm, Witec, GmbH.) with a maximum output of 75 mW was fiber-coupled into the microscope using a 10 µm single mode silica fiber. Raman images were measured using a Leica 10 × /0.25 water immersion objective. The backscattered Raman photons were fed into an imaging spectrograph (UHTS 300, Witec, GmbH.) with a 600 groove/mm grating using a 100 μm silica fiber. The spectrometer was equipped with a thermoelectrically cooled (−60 °C), charge-coupled device camera (Newton, Andor Technology Ltd. UK). The confocal Raman system acquires spectra covering the range from 0 to 3600 cm⁻¹. To calibrate the wavelength axis, the atomic emission lines of the argon/mercury spectral calibration lamp (HG-1, Ocean Optics, Inc.) were used.

Albro M.B., Bergholt M.S., St-Pierre J.P., Vinals Guitart A., Zlotnick H.M., Evita E.G, & Stevens M.M. (2018). Raman spectroscopic imaging for quantification of depth-dependent and local heterogeneities in native and engineered cartilage. NPJ Regenerative Medicine, 3, 3.

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Confocal Raman Spectral Acquisition Methodology

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Confocal Raman spectral acquisition was performed on a Raman micro-spectroscope (alpha300R + , WITec, Ulm, Germany). The light source used was a 785 nm laser (Toptica XTRA II) with a 63 × /1.0 NA water immersion microscope objective lens (W Plan-Apochromat, Zeiss, Oberkochen, Germany). The scattered light was collected via a 100 μm fibre with a 600 groove mm⁻¹ grating spectrograph (UHTS 300, WITec, Ulm, Germany) and spectra were acquired using a thermoelectrically cooled back-illuminated CCD camera (iDus DU401-DD, Andor, Belfast, UK) with a spectral resolution of 3 cm⁻¹ and 85 mW laser power at the sample. Laser control was performed remotely via a serial connection and custom MATLAB (2016b, The Mathworks, MA, USA) scripts.

Penders J., Pence I.J., Horgan C.C., Bergholt M.S., Wood C.S., Najer A., Kauscher U., Nagelkerke A, & Stevens M.M. (2018). Single Particle Automated Raman Trapping Analysis. Nature Communications, 9, 4256.

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Confocal Raman Mapping of Calcifications

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A confocal Raman microscope (alpha300RA, WITec, Ulm, Germany) with a frequency-doubled Nd:YAG laser (532 nm) excitation source was used (maximum power: 75 mW). The charge-coupled device detector (DU401A-BV, Andor, UK) was placed behind the spectrometer (UHTS 300, WITec, Ulm, Germany) with a grating of 600 g/mm (Blaze wavelength = 500 nm). Samples were placed on a multi-axis piezo scanner and a motorized large-area stage for sample positioning and imaged using a water immersion 60× objective [Nikon, numerical aperture (NA) = 1.0]. The lateral resolution was 0.61 λ/NA = ~325 nm. The typical mapping scanning step size used was 1 μm (though ranged from 0.5 to 2 μm) with a typical integration time of 0.23 s per step (though going as high as 4 s per step in rare cases).
Mapping data from 117 regions (from 96 individual calcifications and 5 particle-containing regions) containing Raman-detected calcifications with pathological continuity were analyzed. All Raman data were analyzed using WITec Project Plus version 4.1 and, where indicated, Igor Pro 7 (WaveMetrics Inc., Lake Oswego, OR, USA). Raman data processing is elaborated in Supplementary Text.

Kunitake J.A., Sudilovsky D., Johnson L.M., Loh H.C., Choi S., Morris P.G., Jochelson M.S., Iyengar N.M., Morrow M., Masic A., Fischbach C, & Estroff L.A. (2023). Biomineralogical signatures of breast microcalcifications. Science Advances, 9(8), eade3152.

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Confocal Raman Mapping of PVA Hydrogels

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Each sample was hydrated for more than 2 hours and then pressed between a glass slide and a coverslip to ensure a flat surface. The coverslip was then sealed at the edges with nail polish to prevent the hydrogel from drying. A confocal Raman microscope (alpha300 RA, WITec, Germany) with 20× objective (Zeiss, Germany) was used. An Nd:YAG laser (532 nm) was used as the excitation source with the maximum power of 75 mW. Data were collected using a charge-coupled device detector (DU401A-BV, Andor, UK) behind a grating spectrometer (600 g/mm; UHTS 300, WITec, Germany). A 20-μm-resolution Raman map of 4 × 3–mm scan area was acquired with an accumulation time of 1 s per point. Each point was prebleached for 400 ms to decrease the effect of fluorescence. Cosmic ray removal and background subtraction were performed to clean the spectra. The intensity of O–H bond within the PVA and water was calculated by integrating the spectra in the range of 2800 to 3000 cm⁻¹ and 3075 to 3625 cm⁻¹, respectively. The ratio of PVA and water was then calculated and plotted as a heatmap shown in Fig. 5A.

Lin S., Liu X., Liu J., Yuk H., Loh H.C., Parada G.A., Settens C., Song J., Masic A., McKinley G.H, & Zhao X. (2019). Anti-fatigue-fracture hydrogels. Science Advances, 5(1), eaau8528.

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Raman and IR Analysis of Microsections

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Raman measurements of the microsections were performed with a confocal Raman microscope (Alpha300RA, WITec GmbH, Germany). For exciting the sample, a polarized, coherent compass sapphire VIS laser (λ_ex = 532 nm, WITec GmbH, Germany) was used. The radiation was focused through a 100× oil immersion objective (numerical aperture = 1.4, coverslip correction 0.17 mm) (Carl Zeiss, Germany) onto the sample. Measurements at 785 nm were conducted the same way, but used a linear polarized XTRA II laser (785 nm, Toptica Photonics, Germany). The Raman scattering directed through optical multifiber (50/100 μm diameter) to a spectrometer (UHTS 300 WITec, Germany) (600 g.mm^-1 grating) and finally to the CCD camera (Andor DU401 BV and Andor DU401 DD, Belfast, North Ireland). The lateral resolution was about 0.3 μm one full wavenumber spectrum with an integration time of 0.08 s and a laser power of 35/190 mW (532/785 nm) was obtained from every image pixel. The Control Four (WITec GmbH, Germany) software was used for acquisition of the Raman measurements. Reference spectra of ellagic acid and tannic acid, purchased from Sigma-Aldrich (Vienna, Austria), were measured on the same system.
IR measurements were conducted on an FT-IR ATR spectrometer (Vertex 70, Bruker, Billerica, USA) with 32 scans per measurements. Five measurements were averaged using OPUS 7.5 software from Bruker (Billerica, USA).

Felhofer M., Bock P., Xiao N., Preimesberger C., Lindemann M., Hansmann C, & Gierlinger N. (2021). Oak wood drying: precipitation of crystalline ellagic acid leads to discoloration. Holzforschung, 75(8), 712-720.

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Raman Imaging of Immobilized Vesicles

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To immobilize giant
vesicles, calcium fluoride slides were pretreated with 10 mg/mL protamine
(Protamine sulfate salt from salmon, Sigma) for 10 min. After 5 washes
with DPBS, 20 μL of vesicle suspension was incubated on slides
for 1 h, before 20 washes with DPBS. Vesicles were imaged using the
alpha 300R+ confocal Raman microscope (Witec GmBH, Germany). A 35
mW, 532 nm laser light source was shone through a 63× 1.0 NA
water immersion objective lens (W Plan-Apochromat, Zeiss, Germany).
Raman scattering was collected through the same lens and directed
via a 100 μm diameter silica fiber to a 600 groove/mm spectrograph
(UHTS 300, WITec, GmbH, Germany) coupled to a back-illuminated charge-coupled
device camera, cooled to −60 °C. Area scans of vesicles
were imaged with 500 × 500 XY nm resolution. Spectral preprocessing
was performed with ProjectFIVE software (Witec GmBH). First, cosmic
rays were removed, and then the dark current background was subtracted,
followed by “shape” background correction. Spectra were
normalized to the maximum intensity of the water peak at 3000–3400
cm^–1. Finally, Raman images were reconstructed from
univariate analysis of the intensity of the NH₂ signal at 2208 cm^–1 and the SO₃ signal at 2220 cm^–1. Due to low intensity of the 708 cm^–1 polymer
peak with this instrumental setup, Raman images of polymer signal
were reconstructed from 2905 cm^–1 polymer peak.

Saunders C., Foote J.E., Wojciechowski J.P., Cammack A., Pedersen S.V., Doutch J.J., Barriga H.M., Holme M.N., Penders J., Chami M., Najer A, & Stevens M.M. (2023). Revealing Population Heterogeneity in Vesicle-Based Nanomedicines Using Automated, Single Particle Raman Analysis. ACS Nano, 17(12), 11713-11728.

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SERS Performance Evaluation of Modified Fiber

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BZT (Sigma-Aldrich, USA) was used as the probe molecules to evaluate the SERS performance of the modified fiber. The fiber tip was incubated in 1×10⁻³ M of BZT ethanolic solution for 24h to form a self-assembled monolayer. A confocal Raman microscope (Alpha 300 RSA+, WITec, Germany) equipped with a 785 nm diode laser (Toptica Photonics, Germany) was used for SERS measurement. BZT signals were collected via 20× objective (NA = 0.4) with 3 mW and 0.1 s integration time. Before the measurement, the instrumental calibration was verified using a silicon wafer by the silicon peak at 520 cm⁻¹. The backscattered photons were detected by a spectrometer (UHTS300, WITec, Germany) equipped with a CCD camera (DU401A, Oxford Instruments, UK). Cosmic ray removal and baseline correction were conducted by an instrument embedded software (Project v4.1, WITec, Germany) after the signal acquisition.

Choo P.W., Rand C.S., Inui T.S., Lee M.L., Canning C, & Platt R. (2001). A cohort study of possible risk factors for over-reporting of antihypertensive adherence. BMC Cardiovascular Disorders, 1, 6.

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Raman Imaging of Renal Cortex

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GO deposition in the renal cortex was imaged by a WITec alpha 300 R confocal Raman microscope (WITec, Germany) with a 532 nm laser excitation source. Raman signals were acquired by a Peltier-cooled CCD (−70 °C) detector using a 600 line/mm grating spectrometer (UHTS 300, WITec, Ulm, Germany). Raman images were obtained at 0.5 × 0.5 µm² pixel resolution at 500 ms/point integration time.

Chen W., Wang B., Liang S., Wang M., Zheng L., Xu S., Wang J., Fang H., Yang P, & Feng W. (2022). Understanding the Role of the Lateral Dimensional Property of Graphene Oxide on Its Interactions with Renal Cells. Molecules, 27(22), 7956.

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Raman Microspectroscopy of Skin Lesions

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Micro-Raman spectra were obtained with an Alpha 500R confocal Raman microspectroscopy system (WITec GmbH, Germany) coupled with a helium-neon (He-Ne) continuous 633 nm laser beam (35 mW @ 633 nm, Research Electro-optics, Inc., USA). The excitation laser beam was collimated into a 20× objective lens (NA = 0.85, N-Achroplan, Zeiss, Germany) for Raman excitation. Raman photons were collected by the same objective lens and transmitted through a holographic edge filter to a multi-mode optical fiber (50 μm diameter) to the spectrometer (UHTS300, WITec GmbH, Germany), which was equipped with a resolution about 3 cm⁻¹ over a spectrum range of 0–2400 cm⁻¹. The spectra were recorded using a back-illuminated, deep depletion CCD camera containing 600 × 200 pixels (Du401A-BR-DD-352, Andor Technology, UK) working at −60 °C. The spectral data were acquired point-by-point over each kind of skin lesion with 3 s integration time. Before the experiment, a standard tungsten lamp (RS-3, EG&G Gamma Scientific, USA) was used for calibrating the spectral response of the system, and the Raman spectrum of silicon (520 cm⁻¹) was measured to calibrate the wavelength position.

Zhang X., Yu F., Li J., Song D., Li H., Wang K., He Q, & Wang S. (2019). Investigation on the Cancer Invasion and Metastasis of Skin Squamous Cell Carcinoma by Raman Spectroscopy. Molecules, 24(11), 2059.

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Characterization of CuHCFe Thin Films

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Atomic force microscopy (AFM; Dimension Edge, Bruker, Billerica, MA, USA) was performed to measure the surface morphology of the CuHCFe thin films. Scanning electron microscopy (SEM; Apreo 2, Thermo Fisher Scientific, Waltham, MA, USA) was performed to determine the morphology of the CuHCFe thin films. The ultraviolet–visible (UV–vis) spectra of the K₃Fe(CN)₆ solutions before and after reaction with copper were obtained using the Shimadzu UV 2450 spectrometer. The solutions for UV–vis analysis were contained in disposable cuvettes (UV-cuvette semi-micro, Brand, Wertheim, Germany). The Raman spectrum of the CuHCFe thin film was obtained using a lab-made system with laser (532 nm) and Raman spectrometer (UHTS 300, WITec, Ulm, Germany) at −70 °C.

Yun J., Kim Y., Gao C., Kim M., Lee J.Y., Lee C.H., Bae T.H, & Lee S.W. (2021). Copper Hexacyanoferrate Thin Film Deposition and Its Application to a New Method for Diffusion Coefficient Measurement. Nanomaterials, 11(7), 1860.

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