A piece of the treated ileum described in In vitro intestinal absorption study section was placed on a 22×50 mm coverslip (MENZEL-GLÄSER®, Braunschweig, Germany). Visualization was accomplished using the 10× objective lens system of a Zeiss LSM 510 META inverted microscope (Carl Zeiss, Jena, Germany) equipped with a He–Ne 1 laser (excitation wavelength =543 nm, emission wavelength =580 nm). The tissue was scanned to obtain a series of x–z plane serial images of the Nile red using the laser to scan through the tissue to compare the penetration depths and fluorescence intensity. The fluorescence intensity was determined at the central horizontal line of each image using Zeiss LSM 5 operating software. The mean fluorescence intensity of each image was plotted as a function of the penetration depth.
Lsm 510 meta inverted
Manufactured by Zeiss
Sourced in Germany
The Zeiss LSM 510 META inverted is a confocal laser scanning microscope that uses a unique optical design to provide high-resolution imaging capabilities. It features a multi-track detection system, allowing for simultaneous acquisition of multiple fluorescent signals. The system is designed to deliver precise and accurate data for a variety of imaging applications.
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Lab products found in correlation
Eclipse 80i, by Nikon (2 mentions)
Goat anti mouse fitc, by Jackson ImmunoResearch (1 mentions)
Anti brdu, by BD (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Te2000, by Nikon (1 mentions)
Species specific fluorescently labeled secondary antibodies, by Jackson ImmunoResearch (1 mentions)
Dmi6000b, by Leica (1 mentions)
Apotome 2, by Zeiss (1 mentions)
1x71 inverted microscope, by Olympus (1 mentions)
Tcs sp5 upright confocal microscope, by Leica (1 mentions)
Lsm image browser, by Zeiss (1 mentions)
8 protocols using lsm 510 meta inverted
1
Visualizing Nile Red Penetration in Ileum
2
BrdU Pulse Labeling of Cell Cycle
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MCF10A-MYCER cells were plated on sterilized coverslips. After 24 hr, cells were incubated in 2.5mM thymidine for 17 hr. After a 7 hr recovery, the second block was administered at 2.5mM for 14 hr. Vehicle or 4OHT was added 3 hr prior to release. After 1 hr recovery, cells were treated with an 8 min pulse of 33цM BrdU. Immediately, cells were washed and fixed with 4% paraformaldehyde for 15 min at RT, washed, then permeabilized with 0.2% Triton-X in PBS for 5 min and washed. DNA was denatured in 2N HCl for 20 min at RT, following neutralization with 0.1M sodium tetraborate pH8.5. Primary antibody incubation with anti-BrdU (#555647, BD Pharmingen, San Jose, CA, USA) was performed in PBS-0.5%Tween for 30 min at RT, followed by washing, and secondary antibody incubation with goat-anti-mouse-FITC (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA) was performed in PBS-0.5%Tween for 30 min. Cells were washed and stained for 1 min with propidium iodide, airdried, and mounted on slides for confocal microscopy. The Zeiss LSM 510 Meta Inverted (Carl Zeiss AG, Oberkochen, Germany) was used for all confocal imaging.
3
Visualizing AvrPi9 and PWL2 Localization
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In order to capture the localization of AvrPi9 and it's co-localization with PWL2, AvrPi9-mCherry and PWL2-GFP strains (Table S1) were used for rice leaf sheath assay in the susceptible rice line IR31917, as described previously (Kankanala et al., 2007) . Images were taken at three time points (25, 27 and 32 hpi). Confocal microscopy was performed with Zeiss LSM510 META Inverted (Carl Zeiss Inc., Jena, Germany). The objective was an 963 Achromat (numerical aperture 1.4) oil immersion lens. mCherry fluorescence was imaged with a 543-nm laser and a 560-to 610-nm band-pass emission filter. EGFP fluorescence was imaged with a 488-nm laser and a 505-to 530-nm band-pass emission filter.
4
Retinal Vascular Development in Mice
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Postnatal day 6 (P6), 10 weeks, 4-months, 9 months, and 1-year old mice were sacrificed, enucleated, and the eyes were fixed in 4% paraformaldehyde for 2 h at 4 °C. Retinas were dissected, then incubated for 2 h at room temperature in blocking buffer (PBS, 2% BSA, 0.2% Triton X-100). After three 20 min washes in Pblec buffer (PBS supplemented with 1 mM MgCl2, 1 mM MnCl2, 1 mM CaCl2, and 1% Triton X-100), retinas were incubated overnight at 4 °C with fluorescein labeled isolectin B4 (IB4; Vector Laboratories, FL-1201, 1:25) and antibodies diluted in blocking buffer. Retinas were washed three times with blocking buffer and incubated with species-specific fluorescently labeled secondary antibodies (Jackson Laboratories, 1:100) diluted in blocking buffer for 2 h at room temperature. After three washes in PBS, whole retinas were flat-mounted in ProLong Gold Antifade reagent (Life Technologies) containing Hoechst 33,342 (Life Technologies) and analyzed with an epifluorescence microscope (Nikon TE-2000) or a laser scanning fluorescence microscope (Zeiss LSM 510 Meta inverted).
5
Bright Field and Fluorescence Microscopy Protocol
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Bright field images were acquired using 1X71 inverted microscope (Olympus, Japan), DMI6000B inverted live imaging or TCS SP5 upright confocal microscope (Leica, Germany) at magnifications as indicated. The surface and intracellular fluorescence imaging was performed on APECs stained on glass cover-slips after appropriate treatments. Imaging was performed using the ApoTome.2 inverted fluorescence microscope at 63X or LSM 510 META inverted confocal microscope (Zeiss, Germany) at 100X oil immersion objectives. The images were recorded using the Axio Vision, Kombi-FCS LSM–META or LAS AF softwares. Multiple fields across each sample were acquired.
6
Fluorescent Immunostaining of CD44 in Scaffolds
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Samples were washed in PBS and then they were submerged in 0.1% formaldehyde for 15 minutes. CD44 antibody (Anti-rat with fluorochrome Alexa Fluor 647) (1:250) and DAPI (1:500) in PBS were added to the scaffolds for 1 hour before being washed with PBS. Images (1024 x 1024 pixels) were obtained using a Zeiss LSM 510Meta inverted confocal microscope and x10/0.3 water dipping objective, with a pixel dwell time of 6.4 µs. DAPI was excited using an 780 nm laser (8.1% transmission) and emission detected between 435 and 485 nm. CD44 was excited using a 633 nm laser (51% transmission) and emission detected between 650 and 710 nm. All image analyses were performed using Zeiss LSM image browser and ImageJ.
7
Hippocampal FGF2 and DCX Quantification
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For FGF2 quantification, images covering the whole hippocampal area were captured by a Nikon fluorescent microscope (eclipse 80i) with a mounted digital camera and analyzed using Image J software. Low magnification (4×) images were used to quantify the FGF2 by tracing the area of staining, while the high magnification (20×) images were used for mean pixel intensity quantification using three equal randomly chosen squared areas (100 × 100 pixel or 31.4 × 31.4 μm) and averaged. For DCX immunopositive cell counting, images covering the dentate gyrus were captured by a confocal microscope (Zeiss LSM 510 Inverted Meta) with 20× magnification. DCX positive cells in the subgranular zone (SGZ) were counted in sections through the whole hippocampal formation of each mouse and averaged. Four mice were analyzed for each group.
8
Hippocampal FGF2 and DCX Quantification
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