Lsm 510 meta confocal

Manufactured by Zeiss

Sourced in Germany, United States

The Zeiss LSM 510 META is a confocal laser scanning microscope designed for high-resolution imaging. It utilizes a multi-track detection system with multiple laser sources to enable simultaneous acquisition of multiple fluorescent signals. The instrument provides optical sectioning capabilities for 3D imaging of samples.

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Lab products found in correlation

Guinea pig anti insulin, by Abcam (2 mentions) Sheep anti glucagon, by Merck Group (2 mentions) Rabbit anti somatostatin, by Vector Laboratories (2 mentions) Vectashield, by Vector Laboratories (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Prolong diamond antifade, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Axioskop 2 upright microscope, by Zeiss (1 mentions) Mitotracker red cmxros, by Thermo Fisher Scientific (1 mentions) Plan apochromat 100 1.4 oil objective, by Zeiss (1 mentions) Axio imager m2, by Zeiss (1 mentions) Apotome 2, by Zeiss (1 mentions) Deltavision elite, by GE Healthcare (1 mentions) Orca flash 4.0 lt camera, by Zeiss (1 mentions) Dm6000, by Leica (1 mentions) Lsm 710, by Zeiss (1 mentions) Lsm pascal, by Zeiss (1 mentions) Rabbit anti rfp, by Rockland Immunochemicals (1 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (1 mentions) Cy2 conjugated anti chicken, by Jackson ImmunoResearch (1 mentions)

Spelling variants of this product

LSM 510 (2630 citations) LSM 510 META (1974 citations) 510 Meta (155 citations) Confocal LSM 510 (10 citations) Axiovert 200M confocal LSM 510 META Zeiss microscope (3 citations) LSM 510 Meta Axiovert confocal microscope (3 citations) LSM 510 META Axioplan 2 confocal laser scanning microscope (2 citations) Axio-vision/LSM 510 META inverted confocal microscope (2 citations) LSM 510 Meta NLO multiphoton confocal microscope (2 citations) LSM 510 Meta Confocal Scanning Microscope (2 citations)

31 protocols using lsm 510 meta confocal

FRAP Assay for Fusion Protein Dynamics

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Upon recruitment to anti-ICAM-1-antibody-coated beads, FRAP assays were performed in the area of accumulated fusion protein directly next to the bead. Experiments were performed using 50 iterations with 488-nm laser illumination, at maximum power (25 mW) and a Zeiss LSM-510-META confocal laser scanning microscope. Fluorescence recovery was measured by time lapse imaging for 80s with intervals of 0.784s. The signal was corrected for background and bleaching for each cell and time point. For normalization, the average of the saturated signal was calculated as 100%.

Schaefer A., van Duijn T.J., Majolee J., Burridge K, & Hordijk P.L. (2017). Endothelial CD2AP binds the receptor ICAM-1 to control mechanosignaling, leukocyte adhesion and the route of leukocyte diapedesis in vitro. Journal of immunology (Baltimore, Md. : 1950), 198(12), 4823-4836.

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Immunocytochemical Identification of Islet Cells

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In all recordings, cell identity (α, β or δ) was subsequently established by immunocytochemistry. Biocytin (0.5 mg ml⁻¹) was included in the intracellular solution to allow identification of the cell recorded from. Following voltage-clamp experiments, islets were fixed with 4% formaldehyde in phosphate-buffered saline (PBS) overnight and permeabilized with 0.3% Triton X-100. Non-specific binding was blocked by pre-treatment for 2 h with 5% normal goat serum before incubating with the different primary antibodies for 4–12 h (guinea pig anti-insulin (Abcam, Cambridge, UK), sheep anti-glucagon (Sigma-Aldrich, St Louis, MO) and rabbit anti-somatostatin (Vector Labs, Burlingame, CA)). After washing with PBS, the islet was incubated for 1 h in secondary antibodies (Alexa 633 goat anti-guinea pig (insulin), Alexa 405 goat anti-mouse (glucagon) and Alexa 543 goat anti-rabbit (somatostatin)). Biocytin labelling was visualized by using Alexa Fluor 488 conjugated streptavidin (0.04 mg ml⁻¹; Thermo Fisher). Islets were then washed and imaged on a confocal microscope (Axioskop 2 upright microscope fitted with a Zeiss LSM 510 meta confocal and a chameleon multiphoton module).

Briant L.J., Zhang Q., Vergari E., Kellard J.A., Rodriguez B., Ashcroft F.M, & Rorsman P. (2017). Functional identification of islet cell types by electrophysiological fingerprinting. Journal of the Royal Society Interface, 14(128), 20160999.

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Visualizing Mitochondrial Dynamics in GFP-Tagged Strains

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GFP-tagged strains were cultured to the stationary phase in YPD at 30°C and inoculated into fresh YPD at a density of approximately 1 × 10⁶ cells/ml. After 3 to 4 h of incubation, cells were labeled with MitoTracker Red CMXRos (Molecular Probes), fixed with formaldehyde, and imaged with a Zeiss LSM510 Meta confocal laser scanning microscope equipped with a Plan-Apochromat 100×/1.4 oil objective. Details of the protocol are provided in Text S1.
Colocalization was analyzed as described by Costes et al. (51 (link)) using the Coloc 2 algorithm as implemented in the Fiji package of ImageJ (90 (link)). Images were processed with a 2-pixel median filter, and background was subtracted with a 1-pixel sliding paraboloid before quantitation. A minimum of 30 cells from at least two independent samples were analyzed, and the Costes significance test was based on 100 randomizations.

Sun N., Parrish R.S., Calderone R.A, & Fonzi W.A. (2019). Unique, Diverged, and Conserved Mitochondrial Functions Influencing Candida albicans Respiration. mBio, 10(3), e00300-19.

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Yeast Cell Imaging Protocol

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Yeast samples were cultured in standard SMD medium with 5% glucose (exceptions indicated). Unless noted, images of live cells were collected on a Deltavision Elite deconvolution microscope (GE Healthcare/Applied Precision) on a 100x objective (CoolSnap HQ, Photometrics). 16-image z stacks with 0.25-μm spacing were taken to cover the entire cell. For the experiments to test colocalization, the images of a best-focused single stack were presented. For other purposes, all stacks of images were projected, and analyzed in ImageJ.
Light microscopy images for Supplementary Figures S1C, S2, S5, S6, and 2D were captured on a Zeiss AxioImager M2 with an ORCA-Flash 4.0 LT camera and processed with Zeiss Zen2 software. Representative images of a best-focused single stack are presented. Images for S7 and 4D were captured and processed using a Zeiss Apotome 2.0. For strains in Supplementary Figure S2B with weak GFP signal, a center snap at a longer exposure is used for the representative image. Images for Figure S4 and 2C were imaged on a Zeiss LSM 510 META confocal in z stacks and analyzed using Zeiss Zen 2.0 software.

Jin M., Fuller G.G., Han T., Yao Y., Alessi A.F., Freeberg M.A., Roach N., Moresco J.J., Karnovsky A., Baba M., Yates JR I.I.I., Gitler A.D., Inoki K., Klionsky D.J, & Kim J.K. (2017). Glycolytic Enzymes Coalesce in G bodies Under Hypoxic Stress. Cell reports, 20(4), 895-908.

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Phagocytosis of Opsonized Salmonella by Macrophages

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The semiadherent macrophage-like cell line J774 (24 (link)) was used in this assay, which was performed as described previously (25 (link)). S. Typhimurium–GFP bacteria were opsonized with WT or IgG1ko sera as describe above. Opsonized bacteria were mixed with macrophages in a 5:1 ratio, grown on glass coverslips in serum-free RPMI 1640 medium, and incubated at 37°C for 1 h. Nonadherent bacteria were washed off with PBS 1 h later, and fixed in 4% paraformaldehyde. The coverslips were mounted on microscope slides and visualized on a Zeiss LSM 510 Meta confocal with 100× lens. Fluorescence photomicrographs were taken with a Leica DM6000 with filters suitable for GFP detection. Macrophage cell bodies were segmented using ImageJ/Fiji image analysis software (26 ) by thresholding the autofluorescent signal obtained at 600 ± 20 nm after 546 nm excitation. The percentage of macrophage area covered by GFP was calculated from >1000 cells per sample.

Zhang Y., Dominguez-Medina C., Cumley N.J., Heath J.N., Essex S.J., Bobat S., Schager A., Goodall M., Kracker S., Buckley C.D., May R.C., Kingsley R.A., MacLennan C.A., López-Macías C., Cunningham A.F, & Toellner K.M. (2017). IgG1 Is Required for Optimal Protection after Immunization with the Purified Porin OmpD from Salmonella Typhimurium. The Journal of Immunology Author Choice, 199(12), 4103-4109.

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Neutrophil Adhesion at Endothelial Cell-Cell Borders

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500,000 freshly isolated neutrophils were added to TNFα-treated HUVEC monolayers cultured on a 30 mm glass coverslip and placed in a heating chamber at 37°C and 5% CO₂. After 10 min incubation, non-adherent neutrophils were washed away with EGM-2 medium, cells were fixed and stained with phalloidin (F-actin). Images with three z-stacks (every 2.3 μm) in 14 random fields were captured on a Zeiss LSM510-META confocal microscope (63x/NA 1.4 oil objective) and processed with Zen2009 software and ImageJ. Adherent neutrophils were counted based on DIC images, endothelial cell-cell borders were defined by endogenous F-actin staining (phalloidin). An adherent neutrophil was defined as being located at the cell-cell border when at least half of the neutrophil was adherent above the cell-cell contacts). The percentage of adherent neutrophils above endothelial cell-cell contacts was calculated by the number of bound neutrophils above cell-cell border / number of all adherent neutrophils*100.

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Immunostaining of Adult Fly Brains

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Dissection and immunostaining of adult fly brains was performed as previously described [12 (link)]. The primary antibodies employed were rabbit anti-GFP 1:500 (Invitrogen), rabbit anti-RFP 1:500 (Rockland), chicken anti-GFP 1:500 (Upstate), rabbit anti-PDF 1:1500 (custom-made by NeoMPS, France) and homemade rat anti-Drosophila-PDF 1:500 [12 (link)]. Secondary antibodies used were Cy2-and Cy3-conjugated anti-rabbit, Cy2-conjugated anti-chicken and Cy5- and Cy3-conjugated anti-rat (Jackson ImmunoResearch). Images were taken on either a Zeiss Pascal LSM, a Zeiss LSM 510 Meta confocal or a Zeiss LSM 710 two-photon microscope. After acquisition, images were processed employing Fiji, an ImageJ-based image processing environment [44 (link)].

Gorostiza E.A., Depetris-Chauvin A., Frenkel L., Pírez N, & Ceriani M.F. (2014). Circadian pacemaker neurons change synaptic contacts across the day. Current biology : CB, 24(18), 2161-2167.

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Quantitative Imaging of Mechanosensory Neurons

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For quantitative imaging of Pmec-4-GFP- 3′ UTR ced-7 (sensor) lines, the worms coexpressing Pmec-4-GFP- 3′ UTR ced-7 (sensor) and Pmec-4-mCherry-3′ UTR unc-54 (tbIs222) were imaged with 60.5% of 488-nm and 43.6% of 543-nm lasers, respectively, under a 100× oil objective (NA 1.3). Scanning was done with a Zeiss LSM 510 Meta confocal. Two or three z-sections were collected with a scanning speed of 6 and 2× averaging, with detector gain values of 668 and 708 for the 488-nm and 543-nm lasers, respectively. Images were collected below the saturation limit. The average GFP and mCherry intensities were measured in a ROI drawn on the PLM cell body.
EFF-1::GFP: Pmec-4::EFF-1::GFP was imaged using 68% of a 488-nm laser.

Basu A., Dey S., Puri D., Das Saha N., Sabharwal V., Thyagarajan P., Srivastava P., Koushika S.P, & Ghosh-Roy A. (2017). let-7 miRNA controls CED-7 homotypic adhesion and EFF-1–mediated axonal self-fusion to restore touch sensation following injury. Proceedings of the National Academy of Sciences of the United States of America, 114(47), E10206-E10215.

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Subcellular Localization of Bna.TTG2 Proteins

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To determine the subcellular localization of full-length Bna.TTG2 proteins, 35S:Bna.TTG2s:GFP constructs [fused to green fluorescent protein (GFP)] were introduced into wild-type (WT) Arabidopsis (Col-0). Root tips of 1-week-old T3 homozygous plants were examined under a Nikon Eclipse80i fluorescence microscope first and then imaged under an LSM 510 META confocal microscope (Zeiss).

Li Q., Yin M., Li Y., Fan C., Yang Q., Wu J., Zhang C., Wang H, & Zhou Y. (2015). Expression of Brassica napus TTG2, a regulator of trichome development, increases plant sensitivity to salt stress by suppressing the expression of auxin biosynthesis genes. Journal of Experimental Botany, 66(19), 5821-5836.

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Quantifying Focal Adhesions in Serum-starved Cells

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Serum-starved cells were plated on eight-well slides coated with 1.25 μg/ml FN or 0.1 mg/ml PDL. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.4% Triton X-100, and blocked with 2% BSA in phosphate-buffered saline (PBS). Most cells were stained with rhodamine–phalloidin. Rescue experiments in which DKO cells were infected to express HA-tagged arrestins were also stained with anti-HA antibody to detect expression. Images were taken on a Nikon TE2000-E automated inverted microscope (Nikon, Melville, NY) with 40× oil objective with additional 1.5 optical magnification. Focal adhesion numbers and size were quantified from confocal images taken on LSM 510 Meta Confocal (Zeiss, Jena, Germany) with 40× oil objective and analyzed with ImageJ (National Institutes of Health, Bethesda, MD).

Cleghorn W.M., Branch K.M., Kook S., Arnette C., Bulus N., Zent R., Kaverina I., Gurevich E.V., Weaver A.M, & Gurevich V.V. (2015). Arrestins regulate cell spreading and motility via focal adhesion dynamics. Molecular Biology of the Cell, 26(4), 622-635.

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