Spectra x

Manufactured by Lumencor

Sourced in United States, Japan

The Spectra X is a light engine designed for fluorescence microscopy and other applications. It features six independently controllable LED light sources that can be combined to generate a wide range of wavelength outputs.

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Lab products found in correlation

Eclipse ti, by Nikon (14 mentions) Nis elements software, by Nikon (10 mentions) Flash 4, by Hamamatsu Photonics (9 mentions) Ti e widefield microscope, by Nikon (8 mentions) Na 1.4 oil objective, by Nikon (7 mentions) Perfect focus system, by Nikon (6 mentions) Mitotracker red cmxros, by Thermo Fisher Scientific (6 mentions) No 1.5 glass bottomed dishes, by MatTek (5 mentions) Eclipse ti2 e microscope, by Nikon (4 mentions) Imagexpress micro xl, by Molecular Devices (4 mentions) Axopatch 200b, by Molecular Devices (4 mentions) Orca flash 4, by Hamamatsu Photonics (4 mentions) Nucblue, by Thermo Fisher Scientific (4 mentions) Ixon ultra 897 emccd camera, by Oxford Instruments (3 mentions) Ds qi2 cmos camera, by Nikon (3 mentions) Cfi super plan fluor elwd adm objective lens, by Lumencor (3 mentions) Nis element, by Nikon (3 mentions) Dmi6000b, by Leica (3 mentions) Dmi8 inverted microscope, by Leica (3 mentions) Pclamp 10 software suite, by Molecular Devices (3 mentions)

Close spelling variants of this product

SPECTRA X light engine (114 citations) Spectra X LED light engine (18 citations) Spectra-X light engine LED light (7 citations) Spectra X LED illumination (6 citations) Spectra X LED lamp (6 citations) Spectra X LED system (3 citations) Spectra X LED illumination system (2 citations) Spectra X light (2 citations) Spectra X light engine light source (2 citations) SPECTRA X light engine illumination source (2 citations)

140 protocols using spectra x

Quantitative Fluorescent Microscopy Protocol

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Fluorescent microscopy was performed at the Nikon Imaging Core (UCSD) using a Nikon Eclipse Ti2-E microscope with Plan Apo objectives. Samples were excited by the Lumencor SpectraX and acquired with a DS-Qi2 CMOS camera using NIS-Elements software, or with a laser scanning confocal (A1R HD, Nikon), acquired with an iXon Ultra 897 EMCCD camera (Andor). All AVPV slides were imaged at the same time and under the same conditions. All ARC and thalamic reticular nucleus slides were imaged at the same time and under the same conditions. The number of Kiss1 cells that colocalized with Six3 or atypical signal was determined manually, using FIJI Cell Counter tool. NIS-Elements: General Analysis software was used to objectively quantify the intensity of Kiss1, cFos, and Six3 signals. A signal threshold of 2.5 standard deviations above background was used for defining positive signal of each gene. cFos and Six3 signals were quantified only in Kiss1-positive cells.

Lavalle S.N., Chou T., Hernandez J., Naing N.C., He M.Y., Tonsfeldt K.J, & Mellon P.L. (2022). Deletion of the homeodomain gene Six3 from kisspeptin neurons causes subfertility in female mice. Molecular and cellular endocrinology, 546, 111577.

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Multicolor Epifluorescence Microscopy of Phage-Host Interactions

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Unless otherwise stated, all cell imaging was done on M2G 1.5% agarose pads on a light-emitting diode-based (Lumencor, SpectraX) multicolor epifluorescence microscope consisting of a Leica Dmi8 stand equipped with an immersion oil phase contrast objective (100×, HC PL APO, 1.4 numerical aperture) and an EMCCD camera (Hamamatsu, C9100 02 Cl). Cells were grown overnight in M2G to an OD600 of 0.3–0.4. Images were analyzed and processed using Fiji software (68 (link)). For generating line profiles, the Fiji plugin, MicrobeJ (69 (link)), was used to divide cells into either 26 or 50 bins along the longitudinal axis, and the average fluorescence intensity of each bin was used to generate fluorescence profile plots. The protocol for imaging ϕCbK attachment was adapted from Hinz et al. (15 (link)). Briefly, Sytox Green (ThermoFisher S7020) was added to 1 mL of ϕCbK to a final concentration of 25 µM and allowed to incubate overnight at 4°C. Cells were mixed 1:1 with fluorescently labeled phage and imaged.

Chong T.N., Panjalingam M., Saurabh S, & Shapiro L. (2023). Phosphatase to kinase switch of a critical enzyme contributes to timing of cell differentiation. mBio, 15(1), e02125-23.

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In vivo Calcium Imaging of Trigeminal Neurons

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In vivo calcium imaging of TRNs was performed on a
Leica DMI 4000 B microscopy system (Lumencor Spectra X light engine,
fluorescence cube with beam splitter (Chroma, 59022bs), 63x/1.32 NA oil
objective (Leica), Hamamatsu W-view Gemini Image splitting optics with beam
splitter (Chroma, T570lpxr) and emission filters (Chroma, ET525/50m and
ET632/60m), and a Hamamatsu Orca-Flash 4.0LT digital CMOS camera). Cyan (0.77
mW) and yellow (1.21 mW) illumination from the light engine was used to excite
green TRN::GCaMP6s and red mCherry fluorescence in the TRNs. mCherry
fluorescence was recorded to correct for defocusing artifacts during image
analysis. The emission spectra were split by the image-splitting optics and
projected onto separate parts of the camera chip. To follow calcium transients,
image sequences were recorded at a rate of 10 frames-per-second. To control for
potential habituation to repeated stimuli (Fig.
6) when testing the same neuron ventrally and dorsally with
sequential stimulus protocols, we alternated the sequence of which side was
stimulated first. To map the dorso-ventral TRF, for example, we applied the
first stimulus to the ventral side and the second to the dorsal side and
alternated this sequence in a second data set (Fig. S6). No randomization was
performed in the step, ramp, buzz test.

Nekimken A.L., Fehlauer H., Kim A.A., Manosalvas-Kjono S.N., Ladpli P., Memon F., Gopisetty D., Sanchez V., Goodman M.B., Pruitt B.L, & Krieg M. (2017). Pneumatic stimulation of C. elegans mechanoreceptor neurons in a microfluidic trap. Lab on a chip, 17(6), 1116-1127.

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Mitochondrial Membrane Potential Imaging

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Mitochondrial membrane potential was measured using live-cell imaging following staining with the JC-1 dye (Invitrogen). Cultured control and patient fibroblasts were seeded onto 35 mm MatTek dishes (MatTek, Ashland, MA) at a density of 40,000 cells in complete DMEM. On the day of the assay, the media was removed, and cells were washed with phosphate buffered saline (PBS) containing calcium. One µl of the JC-1 dye, diluted to 1 mg/ml in DMSO, was mixed in 1 ml PBS, added to the dish, and incubated for 30 min at 37 °C, 5% (v/v) CO₂. After incubation, the cells were rinsed with PBS, complete DMEM was added, and the dish was placed into a closed, thermo-controlled (37 °C) stage top incubator above the motorized stage of an inverted Nikon TiE fluorescent microscope equipped with a 60 × optic Nikon, CFI Plan Fluor, NA 1.4 (Nikon Inc. Melville, NY). The JC-1 dye was excited using a diode-pumped light engine (SPECTRA X, Lumencor) and detected using an ORCA-Flash 4.0 sCMOS camera (Hamamatsu) and excitation and emission filters from Chroma Technology Corp (Bellows Falls, VT). Data were collected on approximately 10–20 cells per stage position, with 10–15 stage positions per condition, were analyzed using NIS Elements software (Nikon Inc.). The ratio of red:green (hyperpolarized:depolarized) fluorescence was calculated.

Heiman P., Mohsen A.W., Karunanidhi A., St Croix C., Watkins S., Koppes E., Haas R., Vockley J, & Ghaloul-Gonzalez L. (2022). Mitochondrial dysfunction associated with TANGO2 deficiency. Scientific Reports, 12, 3045.

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Fluorescence Microscopy of Polymer Cells

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The light source for excitation of the polymer was provided by a LED system (Lumencor Spectra X) fibre-coupled to the fluorescence port of the microscope; the illuminated spot on the sample had an area of 0.23 mm². Light powers of the cyan LED used (central wavelength λ = 475 nm) ranged from about 0.8 to 13 mW, measured at the output of the microscope objective (P_obj). Due to optical losses in passing the petri-dish and the substrate, and especially due to the high absorption from the active material, the actual optical power reaching the cells layer, for the maximum case of P_obj = 13 mW (corresponding to an impinging intensity of 57 mW/mm²), was reduced to about P_cell = 0.39 mW (which correspond to an intensity of 1.7 mW/mm²). As for the deposited energy, in the case of 20 ms pulses, the maximum pulse energy density impinging the sample can be calculated as 1.14 mJ/mm², corresponding to 34 μJ/mm² actually reaching the cells.

Martino N., Feyen P., Porro M., Bossio C., Zucchetti E., Ghezzi D., Benfenati F., Lanzani G, & Antognazza M.R. (2015). Photothermal cellular stimulation in functional bio-polymer interfaces. Scientific Reports, 5, 8911.

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Live-cell Imaging of Monolayer Cultures

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Cells were plated at *2.710^5 cells/well in fibronectin-coated (EMD Millipore) 96-well glass-bottom plates (Thermo Scientific or Cellvis) 48 hr before imaging. The following day, monolayers were serum-starved with 0.5% HS, phenol-red-free DMEM/F12 containing 1% Glutamax (Gibco). Monolayers were imaged using a Metamorph-controlled Nikon Eclipse Ti-E epifluorescence microscope with a 20x air objective and a Hamamatsu sCMOS camera. The multi-LED light source SpectraX (Lumencor) and the multiband dichroic mirrors DAPI/FITC/Cy3/Cy5 and CFP/YPF/mCherry (Chroma) were used for illumination and imaging without any spectral overlap. Temperature (37°C), humidity, and CO2 (5%) were maintained throughout all imaging using OKO Labs control units. Time-lapse images were captured at 5-minute intervals. Sample sizes were selected by attempting to capture at least 300 cells from each population. Key conditions from imaging experiments were performed at least twice, with one independent replicate presented in figures. All experiments have at least two technical replicates per independent experimental replicate.

Peterson A.F., Ingram K., Huang E., Parksong J., McKenney C., Bever G.S, & Regot S. (2022). Systematic Analysis of the MAPK Signaling Network Reveals MAP3K Driven Control of Cell Fate. Cell systems, 13(11), 885-894.e4.

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Oxidative Stress in Pericyte Model

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Pericytes were either pre-treated with MG132 (5 µM) for 6 h prior to incubation with α-syn ribbons or fibrils for 24 h. MitoSOX reagent was added to the cells at a final concentration of 5 µM. The cells were subsequently incubated for 30 min at 37 °C. 5% CO₂. To label cell nuclei, the NucBlue Live Cell counterstain was added at the same time as the CellROX reagent. The cells were subsequently imaged using the automated fluorescence microscope ImageXpress Micro XLS (Version 5.3.0.1, Molecular Devices, CA, USA) using the 20 x (0.45 NA) CFI Super Plan Fluor ELWD ADM objective lens and Lumencor Spectra X configurable light engine source. Quantitative analysis of several measures including percentage positive cytoplasmic staining, cytoplasmic intensity measures and granularity measurements were performed using the cell scoring, granularity and integrated morphometry analysis modules on MetaXpress software (Version 5.3.0.1, Molecular Devices, CA, USA). Roughly 500–1000 cells were scored per well with multiple wells (at least three) analysed per sample.

Stevenson T.J., Johnson R.H., Savistchenko J., Rustenhoven J., Woolf Z., Smyth L.C., Murray H.C., Faull R.L., Correia J., Schweder P., Heppner P., Turner C., Melki R., Dieriks B.V., Curtis M.A, & Dragunow M. (2022). Pericytes take up and degrade α-synuclein but succumb to apoptosis under cellular stress. Scientific Reports, 12, 17314.

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Immunofluorescence Analysis of CD47 in HEK 293 Cells

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For the immunofluorescence analysis, HEK 293 cells were transfected with either control siRNA or siRNAs targeting HNRNPC as described under Cell Culture and RNAi, 48 h post transfection cells were fixed with 4% paraformaldehyde for 30 min, permeabilized, and blocked with PBS containing 1% BSA and 0.1% Triton X-100 for 30 min. Primary anti-CD47 antibody (sc-59079 from Santa Cruz Biotechnology) was incubated for 2 h at room temperature at a dilution of 1:100 in the same buffer. To visualize CD47 in cells, secondary antibody conjugated with Alexa Fluor 488 was applied, while the nucleus was labeled with Hoechst dye. Imaging was performed with a Nikon Ti-E inverted microscope adapted with a LWD condenser (WD 30mm; NA 0.52), Lumencor SpectraX light engine for fluorescence excitation LED transmitted light source. Cells were visualized with a CFI Plan Apochromat DM 60× lambda oil (NA 1.4) objective, and images were captured with a Hamatsu Orca-Flash 4.0 CMOS camera. Image analysis and edge detection was performed with NIKON NIS Elements software version 4.0. All images were subsequently adjusted uniformly and cropped using Adobe Photoshop CS5.

Gruber A.J., Schmidt R., Gruber A.R., Martin G., Ghosh S., Belmadani M., Keller W, & Zavolan M. (2016). A comprehensive analysis of 3′ end sequencing data sets reveals novel polyadenylation signals and the repressive role of heterogeneous ribonucleoprotein C on cleavage and polyadenylation. Genome Research, 26(8), 1145-1159.

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Microfluidic Imaging Techniques

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Fluids were actuated using both a syringe pump (neMESYS, Cetoni) and/or a pressure pump (OB-1, Fluigent), depending on the applications. Polytetrafluoroethylene (PTFE) tubing was used for fluidic connections. A high-speed camera (EoSens 4-CXP, Mikrotron) was used for imaging on an inverted microscope (Axio Observer Z1, Carl Zeiss) or a stereo microscope (Stemi 2000, Carl Zeiss). Fluorescence imaging was performed with a PCO.edge (PCO) camera and a Spectra-X (Lumencor) light source on the inverted microscope.

Tovar M., Weber T., Hengoju S., Lovera A., Munser A.S., Shvydkiv O, & Roth M. (2018). 3D-glass molds for facile production of complex droplet microfluidic chips. Biomicrofluidics, 12(2), 024115.

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Time-lapse fluorescence microscopy protocol

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Fluorescence and time-lapse microscopy was performed with an inverted fluorescence microscope (Nikon Ti-E, Nikon) equipped with an incubation chamber, an Orca Flash 4 digital camera [Hammamatsu a pE-100-LED (CoolLED)] as the transmission-light source, a Spectra X (Lumencor) as the fluorescent-light source and a 10× objective (Plan Apo λ; numerical aperture, 0.45; DIC N1; working distance, 4). Bright-field images (3% intensity, 90-ms exposure), d2EYFP fluorescence images (excitation, 513/17 nm; intensity, 50%; exposure, 200 ms; YFP ET filter, dichroic 520 nm; emission, 543/22 nm), and mCherry fluorescence images (excitation, 549/15 nm; intensity, 50%; exposure, 200 ms; CY3 HC, dichroic 562 nm; emission, 593/40) were collected. A binning of 2 × 2 was used.

Kim H., Bojar D, & Fussenegger M. (2019). A CRISPR/Cas9-based central processing unit to program complex logic computation in human cells. Proceedings of the National Academy of Sciences of the United States of America, 116(15), 7214-7219.

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