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Lsm 510 meta

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The LSM 510 META is a confocal laser scanning microscope designed for versatile imaging applications. It features a multi-track beam path configuration, allowing for simultaneous detection of multiple fluorescent labels. The system utilizes an array of laser lines and spectral detectors to enable flexible and efficient image acquisition.

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Lab products found in correlation

Tcs sp5, by Leica (2 mentions) Cellmask orange plasma membrane stain, by Thermo Fisher Scientific (2 mentions) Tcs sp8 confocal microscope, by Leica (2 mentions) Alexa fluor 555 goat anti mouse, by Thermo Fisher Scientific (2 mentions) Antibody against tau12, by Abcam (2 mentions) Axioplan 2 microscope, by Zeiss (2 mentions) Lsm 710 quasar, by Zeiss (2 mentions) Illustrator cs4, by Adobe (2 mentions) Sp8 confocal imaging system, by Leica (2 mentions) Lsm image browser software, by Adobe (1 mentions) E600 microscope, by Nikon (1 mentions) Lsm710, by Leica (1 mentions) Lsm780, by Leica (1 mentions) Glass bottomed dishes, by MatTek (1 mentions) Volocity, by PerkinElmer (1 mentions) Transit lt1, by Mirus Bio (1 mentions) Lsm 780, by Zeiss (1 mentions) Axio imager z1, by Zeiss (1 mentions) Lsm image browser, by Zeiss (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

LSM 510 (36 citations) 510 META (3 citations) LSM 510-Meta confocal microscope (3 citations) LSM 510-META Laser Scanning Confocal Microscope (2 citations)

22 protocols using lsm 510 meta

1

Quantifying Melanopsin Dendritic Complexity

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Epifluorescence images were acquired on a Nikon E600 microscope equipped with a SPOT-RT Slider digital microscope camera. For wholemounts, we collected z-stacks of confocal images separated in depth by 0.5 or 1.0 µm on a Zeiss LSM510 Meta or Leica TCS SP5 confocal laser scanning microscope. Confocal images were analyzed using Zeiss LSM Image Browser software and further processed with Adobe Photoshop and ImageJ software.
To assess the density of outer retinal dendrites, we selected 3–5 regions of interest (680 µm × 450 µm) in wholemount retinas from each age studied (P4, P8, P12 and adult). In each of these regions, we counted the number of outer retinal dendrites (ORDs), defined as melanopsin immunopositive dendrites that extended outward into the inner plexiform layer. These dendrites were subdivided into simple and complex varieties based on whether, after entering the INL, they terminated within it (simple) or extended into and coursed within the OPL (complex).
Renna J.M., Chellappa D.K., Ross C.L., Stabio M.E, & Berson D.M. (2015). Melanopsin ganglion cells extend dendrites into the outer retina during early postnatal development. Developmental neurobiology, 75(9), 935-946.
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2

Live Imaging of Mouse Embryo Development

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For live imaging, embryos were cultured in glass-bottomed dishes (MatTek) in an environmental chamber as previously (Kang et al., 2013 (link)). Live imaging conditions used were compatible with normal development as shown previously (Plusa et al., 2008 (link)). For incubation experiments an ERK1/2 inhibitor, 1 μM PD0325901 (StemGent) was added to medium 2-3 hours prior to initiation of 3D time-lapse imaging. Green fluorescent protein (GFP) was excited using a 488-nm Argon laser. Live image data were acquired using three laser scanning confocal imaging systems: Zeiss LSM510META, LSM710, LSM780 and Leica SP8. Images were acquired using 20×/0.75, 40×/1.3 or 63×/1.4 objectives. 20-30 xy planes separated by 2 μm were acquired per z-stack, every 15 minutes. Movies of time-lapse sequences were compiled and annotated using QuickTime Pro (Apple Inc.).
Xenopoulos P., Kang M., Puliafito A., Di Talia S, & Hadjantonakis A.K. (2015). Heterogeneities in Nanog expression drive stable commitment to pluripotency in the mouse blastocyst. Cell reports, 10(9), 1508-1520.
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3

PSCA-Targeted Delivery of TL3 Agonist (dsRNA)

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Example 15

Analysis of Receptor-Mediated Endocytosis of PSCA-Specific Immunoconjugates for Targeted Delivery of TL3 Agonist (dsRNA)

To demonstrate RIBOXXOL® uptake via PSCA receptor-mediated endocytosis the RIBOXXOL® dsRNA was labeled with mal20-PPI-FITC at molar ration of 1:2. For the experiment 6×105 293TPSCA cells grown on a cover slip were treated with FITC-labeled BICs containing RIBOXXOL® and scFv(h-AM1)-BAP at 37° C. in a humidified CO2 incubator, to visualize internalized BICs. As control 293TPSCA cells were treated with FITC-labeled BICs containing RIBOXXOL® and scFv(MR1.1)-BAP, which do not bind to 293TPSCA cells. After 24 h, cell membranes and nuclei were stained with Texas Red®-X conjugate of Wheat germ agglutinin (WGA) and Hoechst (Invitrogen, Waltham, USA) according to the protocols of the manufacturers. Subsequently, the slides were cover slipped in a drop of mounting medium (Vector Laboratories, CA, USA) and examined by a confocal laser scanning microscope (LSM 510 Meta, Leica, Wetzlar, Germany).

FIG. 17 shows the targeted delivery of scFv(h-AM1)-BAP guided BICs containing RIBOXXOL® to 293TPSCA cells. FITC-signals for RIBOXXOL® (see arrows) are only seen in 293TPSCA cells treated with scFv(h-AM1)-BAP containing immunoconjugates whereas o signals for FITC-labeled RIBOXXOL® is detected in cells treated with an EGFRvIII-specific immunoconjugate.

US11504432B2. Delivery system for targeted delivery of a therapeutically active payload (2022-11-22). TECHNISCHE UNIVERSITÄT DRESDEN [DE]. Inventors: Achim Temme [DE].
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4

Quantifying NRF2 and p62 bodies in HeLa cells

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HeLa cells were cultured in 8-chambered cover slides (Nunc) and transiently transfected with 20 nM siRNA, or 50 ng pDestEGFP-NRF2 and 150 ng pDestCherry-SPBP using TransIT-LT1 (Mirus Bio) according to the manufacturer's protocol. Live cell images were taken one day post transfection using a Leica SP5 or a LSM510-META confocal laser scanning microscope. For quantification of p62 bodies, siRNA transfected cells were fixed in 4% paraformaldehyde 2 days post transfection. The cells were permeabilised by 0.1% Triton X-100, for 5 min at room temperature. Staining with antibodies was as described previously [32] (link). Images were obtained using a LSM510-META confocal microscope and p62 bodies were counted manually. Pearson's colocalisation coefficient and scatter were generated using Volocity (Perkin Elmer). Images were processed using Canvas version 11 (ACD systems).
Darvekar S.R., Elvenes J., Brenne H.B., Johansen T, & Sjøttem E. (2014). SPBP Is a Sulforaphane Induced Transcriptional Coactivator of NRF2 Regulating Expression of the Autophagy Receptor p62/SQSTM1. PLoS ONE, 9(1), e85262.
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5

Immunostaining and FISH Imaging Protocol

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Whole-mount and paraffin section immunostaining was performed as previously described32 (link). Intestinal villus images from Figs. 1f and 4h had epithelial cells removed prior to imaging. Primary antibodies are listed in Supplementary Table 1 and were resuspended according to manufacturers’ recommendations when supplied lyophilized. Alexa Fluor 488, 555, and 647 fluorochrome-conjugated secondary antibodies (Invitrogen) were used for signal detection. Nuclei were detected with DAPI. For FISH, probes were purchased from RNAscope (Mm-Lgr5-C3; Mm-Vegfa-ver2-C3; Mm-Adamts18) and hybridization was performed according to manufacturer’s recommendations. Confocal images were obtained using Zeiss LSM 780, Zeiss LSM 510 META or Leica SP5 TANDEM microscopes and standard fluorescent images were obtained using a Zeiss Axio Imager Z1. Images were analyzed using Imaris (Bitplane), ImageJ (NIH) and Photoshop (Adobe) software.
Bernier-Latmani J., Mauri C., Marcone R., Renevey F., Durot S., He L., Vanlandewijck M., Maclachlan C., Davanture S., Zamboni N., Knott G.W., Luther S.A., Betsholtz C., Delorenzi M., Brisken C, & Petrova T.V. (2022). ADAMTS18+ villus tip telocytes maintain a polarized VEGFA signaling domain and fenestrations in nutrient-absorbing intestinal blood vessels. Nature Communications, 13, 3983.
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6

Immunofluorescence Imaging of Tau Protein

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Immunofluorescence confocal microscopy was carried out using Zeiss LSM 510 Meta and Leica TCS SP8 Confocal microscope. Antibody against Tau12 (Abcam, 1:200) and secondary antibody goat anti-mouse Alexa Fluor 555 (Invitrogen, 1:500) were used. CellMask Orange Plasma Membrane Stain (Thermo Fisher Scientific) was used at 0.5x dilution to label the plasma membrane.
Jiang S., Maphis N.M., Binder J., Chisholm D., Weston L., Duran W., Peterson C., Zimmerman A., Mandell M.A., Jett S.D., Bigio E., Geula C., Mellios N., Weick J.P., Rosenberg G.A., Latz E., Heneka M.T, & Bhaskar K. (2021). Proteopathic tau primes and activates interleukin-1β via myeloid-cell-specific MyD88- and NLRP3-ASC-inflammasome pathway. Cell reports, 36(12), 109720.
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7

Polymersome Uptake in Epithelial Cells

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Polymersomes were added at a concentration of 1 mg/ml into the apical (upper) transwell compartment after Trans-Epithelial Electric Resistance (TEER) measurements were taken with an EVOM2 (link) Epithelial Voltohmmeter. For the initial uptake experiments, cells were incubated for 3–6 hours at 37 °C in 95% air 5% CO2, followed by fixation using 3.7% formaldehyde. Where immunofluorescence was performed, fixation was followed by a 30-minute incubation in 0.3% Triton X-100 and 1% bovine serum albumin (BSA). The transwell insert membrane was excised using a scalpel, and mounted on glass cover slip with VectaShield mounting medium. Cells were imaged on a ZEISS LSM 510 META confocal laser-scanning microscope and Leica SP8 confocal laser-scanning microscope with 40x water immersion lens and 63x oil immersion lens. For rhodamine-labelled polymersomes, an excitation energy 561 nm was used and fluorescence emission was measured at 575–600 nm. Nuclear staining was performed using Hoechst 33342 (500 nM) for 10 min in PBS. Image data was acquired and processed using Zeiss LSM Image Browser, Zeiss LSM Image Expert, Leica and Image J software. The acquisition of co-localisation data by means of Pearson’s correlation coefficient was done via the ImageJ plug-in ‘Colocalization Finder’.
Tian X., Nyberg S., S. Sharp P., Madsen J., Daneshpour N., Armes S.P., Berwick J., Azzouz M., Shaw P., Abbott N.J, & Battaglia G. (2015). LRP-1-mediated intracellular antibody delivery to the Central Nervous System. Scientific Reports, 5, 11990.
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8

Visualizing SR-A Expression in Cultured Cells

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Cells were harvested into cover-glass chambers and then cultured in DMEM with 10% bovine serum for 37°C and 5% CO2 for 24 h. Unattached cells were removed by gentle medium exchange, and the attached cells were washed twice with PBS. Cells were fixed with 4% paraformaldehyde. Next, the cells were incubated for 30 min at room temperature with rabbit anti-SR-A antibody (1:100; Abcam), then incubated with FITC-conjugated goat anti-rabbit IgG antibody (1:500; Proteintech Group Inc., Rosemont, IL, USA) for 30 min at 37°C. After staining with DAPI (1:1,000; Thermo Fisher Scientific, Waltham, MA, USA), cells were observed with Laser Scanning Confocal Microscope (LSM-510 Meta; Leica, Frankfurt, Germany).
Xie L., Li Q., Dong R., Zhao K., Feng Y., Bao Z, & Zhou M. (2018). Critical regulation of inflammation via class A scavenger receptor. International Journal of Chronic Obstructive Pulmonary Disease, 13, 1145-1155.
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9

Immunohistochemical Analysis of Drosophila Brains

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Brains were dissected in cold PBS 1X pH 7.4, fixed in 4% formaldehyde/PBT (PBS with 0.1% Triton X-100) for 25 minutes, then washed three times 15 minutes in PBT, and incubated overnight at 4°C with 0.3% Triton X-100 and 1% BSA. Samples were then incubated overnight at 4°C with primary antibodies in PBT, washed in PBT, and incubated overnight with secondary antibodies (4°C). After three washes in PBT, samples were mounted for analysis with Zeiss LSM 510 META or Leica SPE confocal microscopes. The following primary antibodies were used: mouse anti-FasII antibody (1:15; mAb1D4 from Developmental Studies Hybridoma Bank); rabbit anti-Hrp48 antibody (1:400; gift from D. Rio); rabbit anti-GFP antibody (1:1000; A1222, Molecular Probes). Cy3- or Cy5-coupled secondary antibodies (Jackson) were used at 1:1000.
Bruckert H., Marchetti G., Ramialison M, & Besse F. (2015). Drosophila Hrp48 Is Required for Mushroom Body Axon Growth, Branching and Guidance. PLoS ONE, 10(8), e0136610.
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10

Immunofluorescent Localization of UBF

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Cells were seeded on coverslips, washed with PBS, and fixed by incubation in 4% formaldehyde for 20 min at room temperature. The cells were then washed twice in PBS and permeabilized in PBS containing 0.2% Triton x100 and 1% BSA at room temperature for 30 min. Cells were probed with primary antibody to UBF (sc-13125, Santa Cruz), and conjugated secondary anti-mouse Alexa Flour 647 (cat No. 705-605-147, Jackson Labs) antibodies were then applied for detection. To stain the nuclei, the cells were incubated for 30 min with 0.05 μg/ml Hoechst dye (Sigma). Cells were examined and photographed under a confocal laser-scanning microscope (Zeiss LSM 510 META, or Leica STED Live Imaging).
Gelgor A., Gam ze Letova C., Yegorov Y., Kalt I, & Sarid R. (2018). Nucleolar stress enhances lytic reactivation of the Kaposi’s sarcoma-associated herpesvirus. Oncotarget, 9(17), 13822-13833.
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