Tcs nt confocal microscope

Manufactured by Leica

Sourced in Germany

The TCS NT confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a scanning laser unit that enables optical sectioning and high-resolution imaging of specimens. The microscope provides a versatile platform for researchers and scientists to study a wide range of sample types with enhanced contrast and resolution.

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Lab products found in correlation

Lsc lite version 2, by Leica (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Mowiol, by Merck Group (2 mentions) Bx61 fluorescence microscope, by Olympus (2 mentions) Prolong antifade reagent, by Thermo Fisher Scientific (2 mentions) Ms medium, by Duchefa Biochemie (1 mentions) Argon 488 nm, by Leica (1 mentions) Hene 633 nm lasers, by Leica (1 mentions) Plan fluo 40 oil objective, by Nikon (1 mentions) Fv1000 confocal microscope, by Olympus (1 mentions) Dmrbe microscope, by Leica (1 mentions) Dmi6000 microscope, by Leica (1 mentions) Sp5 aobs confocal microscope, by Leica (1 mentions) Axiophot microscope, by Zeiss (1 mentions) Spot software, by Visitron (1 mentions) Vectashield mounting medium, by Vector Laboratories (1 mentions) Anti mouse alexa fluor 555, by Thermo Fisher Scientific (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) Tween 20, by Bio-Rad (1 mentions) Triton x, by Merck Group (1 mentions)

Spelling variants of this product

Confocal microscope (1146 citations) TCS-NT (62 citations) TCS/NT confocal laser scanning microscope (9 citations) TCS-NT SP2 laser confocal microscope (5 citations) TCS NT confocal laser microscope (4 citations) TCS-NT 4D confocal microscope (4 citations) Leica TCS-NT/SP confocal microscope (3 citations) TCS NT upright confocal microscope (2 citations)

24 protocols using tcs nt confocal microscope

Protoplast Transformation for SlCES-YFP Localization

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Protoplast were generated from leaves of 30-day-old plants, grown under sterile conditions on 1/2 MS medium (Duchefa, Haarlem, NL) solidified with 0.7% w/v agar (Duchefa, Haarlem, NL). Protoplasting of mesophyll cells was performed from leaf strips, with a cellulase-based cell wall digestion. The protoplasts were then transformed with the 35S:SlCES-YFP wild-type and mutant constructs, with a polyethyleneglycol-mediated transformation protocol, using the same methodology as described previously for A. thaliana (Khan et al., 2014 (link)).
Following protoplast transformation, SlCES-YFP wild-type and mutant localization was analyzed with or without treatment with 1 μM 24-epiBL or bikinin for 2 h, (as described previously in Khan et al., 2014 (link)), with a Leica TCS-NT confocal microscope (Leica, Weltzlar, Germany). YFP-tagged fusion proteins were excited using the Ar laser line at 514 nm and detected at 520–550 nm. The images were assembled with the Leica confocal software LSC Lite version 2.61.

Shuai H., Chen T., Wlk T., Rozhon W., Pimenta Lange M.J., Sieberer T., Lange T, & Poppenberger B. (2022). SlCESTA Is a Brassinosteroid-Regulated bHLH Transcription Factor of Tomato That Promotes Chilling Tolerance and Fruit Growth When Over-Expressed. Frontiers in Plant Science, 13, 930805.

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GFP-EcO157 Internalization in Tetrahymena

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Internalization of GFP-EcO157 (MM123) in Tetrahymena was monitored after sorting using a Leica TCS-NT confocal microscope equipped HC PL FLUOTAR 20×/0.50 or 40×/0.70 or 63×/1.2 W PL APO objectives, with argon (488 nm), krypton (568 nm), and He/Ne (633 nm) lasers (Leica Microsystems, Wetzlar, Germany) and using Leica TCS NT Software (v. 2.5) for image analysis and preparation. The BP520/50 emission filter set with 488nm laser excitation was used for visualizing the GFP fluorescence. The GFP fluorescence was assigned a green color in the compiled images.

Hernlem B.J., Ravva S.V, & Sarreal C.Z. (2014). Rapid detection of predation of Escherichia coli O157:H7 and sorting of bacterivorous Tetrahymena by flow cytometry. Frontiers in Cellular and Infection Microbiology, 4, 57.

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Imaging Embryos and Ovaries

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Embryos and dissected ovaries were imaged according to a previously described protocol (Dong et al., 2015 (link); Huang et al., 2011 (link)). Embryos were collected overnight at 25°C. Ovaries from adult females several days old were dissected in halocarbon oil (#95). Dechorinated embryos or dissected ovaries were mounted in halocarbon oil on an air-permeable membrane (YSI Membrane Model 5793; YSI) sealed by vacuum grease on a custom-made plastic slide over a 10 × 10–mm cut-through window. After placing the coverslip on top, a membrane at the bottom ensures sufficient air exchange to samples during the imaging session. The slide was then mounted in an air-tight microchamber (custom made) for live imaging under a confocal microscope. Oxygen levels inside the chamber were controlled by flow of either air or custom O₂/N₂ gas at the rate of ∼1–5 cc/s. Images were captured at room temperature (25°C) on a Leica TCS-NT confocal microscope (PL APO 40× oil objective, NA 1.25) by Leica TCS-NT software; an Olympus FV1000 confocal microscope (40× Uplan FL N oil objective, NA 1.3) by Olympus FV10-ASW software; or a Nikon A1 confocal microscope (Plan Fluo 40′ oil objective, NA 1.3) by NIS-Elements AR software. Images were further processed in ImageJ (National Institutes of Health) and Adobe Photoshop.

Dong W., Lu J., Zhang X., Wu Y., Lettieri K., Hammond G.R, & Hong Y. (2020). A polybasic domain in aPKC mediates Par6-dependent control of membrane targeting and kinase activity. The Journal of Cell Biology, 219(7), e201903031.

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Confocal Imaging and Morphometric Analysis

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Embryos in 70% glycerol were mounted on glass slides. Fluorescent images were obtained using a Leica TCS-NT confocal microscope (Leica, Heerbrugg, Switzerland) attached to a Leica DM RBE microscope or a Leica SP5-AOBS confocal microscope attached to a Leica DM I6000 microscope. Confocal images were processed using ImageJ (NIH, Bethesda, MD, USA). Bright-field images were obtained under a Zeiss Axiophot microscope (Zeiss, Munich, Germany). All images were assembled with Adobe Photoshop. All panels are lateral views with the anterior towards the left and dorsal towards the top, unless indicated otherwise.
Cell counting and the measurement of length or area were carried out either manually under microscope or with ImageJ. For example, we manually counted the hair cell number in representative cross sections of the utricle and saccule aided by myo6 and Phalloidin staining. The measurement of the largest diameter of the otocyst and the largest area of the statoacoustic ganglion (SAG) on axial cross sections were performed using ImageJ.

Sun L., Ping L., Gao R., Zhang B, & Chen X. (2023). lmo4a Contributes to Zebrafish Inner Ear and Vestibular Development via Regulation of the Bmp Pathway. Genes, 14(7), 1371.

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Immunofluorescence Localization of HA-Tagged Proteins

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Huh7 cells were grown on glass coverslips and fixed with 3% paraformaldehyde in PBS at RT for 30 min. Cells were blocked and permeabilized with 0.5% BSA and 0.1% Triton X-100 in PBS at RT for 30 min. For localization studies in transfected cells, mouse anti-HA1.1 Epitope Tag (BioLegend, San Diego, CA, USA) was diluted 1:500 into 1% BSA in PBS and incubated at RT for 60 min. Secondary FITC-labeled anti-mouse antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) were used according to the manufacturers’ instructions. After washing, coverslips were mounted on Mowiol (Sigma-Aldrich, St. Louis, MO, USA) and images were taken with a Leica TCS NT confocal microscope (Leica Microsystems, Wetzlar, Germany).

Lundberg R., Melén K., Westenius V., Jiang M., Österlund P., Khan H., Vapalahti O., Julkunen I, & Kakkola L. (2019). Zika Virus Non-Structural Protein NS5 Inhibits the RIG-I Pathway and Interferon Lambda 1 Promoter Activation by Targeting IKK Epsilon. Viruses, 11(11), 1024.

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Microscopy Imaging Protocol

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We used a Nikon H800 microscope (Nikon, Germany) with a SPOT FLEX 64 Mp Shifting Pixel CCD-camera (model #15.2, Diagnostic Instruments Inc., Visitron Systems GmbH, Germany) and SPOT software (version 4.6, Visitron Systems), as well as a Leica TCS NT confocal microscope with Leica confocal software (version 2.00, build 0871, Leica Microsystems, Germany).

Deres F., Schwartz S., Kappes-Horn K., Kornblum C, & Reimann J. (2021). Early Changes in Skeletal Muscle of Young C22 Mice, A Model of Charcot-Marie-Tooth 1A. Journal of Neuromuscular Diseases, 8(Suppl 2), S283-S299.

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Immunofluorescence Analysis of p50 in Macrophages

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PMA-differentiated THP-1 macrophages or serum-differentiated primary human macrophages were grown on glass coverslips before incubation with or without adenovirus. The culture medium was replaced and cells treated as stated in the figure legends. The cells were then washed three times with PBS, fixed and permeabilised with 4% (w/v) paraformaldehyde in PBS at room temperature for 15 min and 100% methanol at −20 °C for 10 min. They were washed twice with PBS containing 1% (w/v) BSA and 0.01% sodium azide. The cells were then incubated with goat polyclonal anti-p50 IgG (5 μg/ml) for 1 h at room temperature, washed four times with PBS containing 1% (w/v) BSA and 0.01% sodium azide and incubated with FITC-or Cy3-conjugated donkey anti-goat IgG (5 μg/ml) and propidium iodide for 1 h at room temperature. Coverslips were then mounted on the slides using VECTASHIELD™ mounting medium (Vector Laboratories, U.K.) and sealed with nail varnish before storage at 4 °C in the dark. Microscope slides were analysed using a Leica TCS NT confocal microscope (Leica Microsystems) using a 63× immersion objective lens. Semi-quantification was performed using Leica confocal software.

Gerry A.B, & Leake D.S. (2014). Effect of low extracellular pH on NF-κB activation in macrophages. Atherosclerosis, 233(2), 537-544.

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Immunofluorescence Imaging of Imaginal Discs

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Third instar imaginal discs were fixed and stained by standard procedures using 4D4 anti-Wg antibody (1:100 dilution, Developmental Studies Hybridoma Bank), secondary antibody anti-mouse-AlexaFluor-555 (1:200 Molecular Probes) and DAPI (Sigma Aldrich). Discs were mounted in Fluoromount G (Southern Biotechnology) and the images were captured on a Leica TCS-NT Confocal microscope.
Fluorescence microscopy of transfected HeLa cells was performed essentially as previously described (10 (link),52 (link)).

Giaimo B.D., Ferrante F., Vallejo D.M., Hein K., Gutierrez-Perez I., Nist A., Stiewe T., Mittler G., Herold S., Zimmermann T., Bartkuhn M., Schwarz P., Oswald F., Dominguez M, & Borggrefe T. (2018). Histone variant H2A.Z deposition and acetylation directs the canonical Notch signaling response. Nucleic Acids Research, 46(16), 8197-8215.

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Particle Internalization in N27 Cells

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N27 cells (15,000/well) were plated on PDL-coated coverslips and particles were added. After 24 hours, cells were washed several times with sterile PBS and fixed in 4% PFA. Non-specific binding was blocked using 2% bovine serum albumin (BSA, (Sigma) in PBS with 0.1% Triton-X (Sigma) and 0.01% Tween-20 (Bio-Rad) for 30 minutes. Cells were stained for 20 minutes with 10 μM phalloidin (Invitrogen). Nuclei were stained with 2 μg/mL Hoechst 33342. Images were taken using an inverted Leica TCS NT confocal microscope.

Brenza T.M., Ghaisas S., Vela Ramirez J.E., Harischandra D., Anantharam V., Kalyanaraman B., Kanthasamy A.G, & Narasimhan B. (2016). Neuronal Protection against Oxidative Insult by Polyanhydride Nanoparticle-based Mitochondria-targeted Antioxidant Therapy. Nanomedicine : nanotechnology, biology, and medicine, 13(3), 809-820.

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Visualizing Influenza Virus Protein Localization

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A549 cells grown on glass coverslips were infected with the wt or NES-mutant viruses at a multiplicity of infection (MOI) -value of 1 and incubated at 37°C 5% CO₂ for 1 h, after which the virus inoculum was removed and replaced with MEM containing 2% FCS. Incubation was resumed and at 8 h, 12 h, 16 h, 20 h and 24 h post-infection infected cells were fixed with 3% paraformaldehyde at room temperature for 20 min, permeabilized with 0,1% Triton X-100 for 5 min and processed for immunofluorescence microscopy as described previously [48 (link)]. The localization of NS1 and NP in the cells were visualized with anti-NS1 and anti-NP antibodies (1:100 dilution) and photographed on a Leica TCS NT confocal microscope. For the leptomycin B (LMB) experiment 2 ng/ml of LMB was added on A549 cells infected with wt virus at 3 h post-infection and the cells were fixed at 20 h post-infection and processed as described above.

Tynell J., Melén K, & Julkunen I. (2014). Mutations within the conserved NS1 nuclear export signal lead to inhibition of influenza A virus replication. Virology Journal, 11, 128.

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