All cells were grown and treated on microslides. Cells were fixed with 4% paraformaldehyde, and confocal images were obtained with a Zeiss LSM 780 Inverted confocal/multiphoton microscope.
Lsm780 inverted
Manufactured by Zeiss
Sourced in Germany
The LSM780 inverted is a laser scanning microscope system designed for high-resolution imaging of biological samples. It features an inverted optical configuration, providing easy access to the specimen and enabling live-cell experiments. The system utilizes a range of laser sources and sensitive detectors to capture detailed images of fluorescently labeled samples. The core function of the LSM780 is to enable researchers to obtain high-quality, confocal microscopy data for their scientific investigations.
Automatically generated - may contain errors
Lab products found in correlation
Zen software, by Zeiss (1 mentions)
Bovine growth serum (bgs), by Thermo Fisher Scientific (1 mentions)
Alexa 647, by Thermo Fisher Scientific (1 mentions)
Actc1, by Merck Group (1 mentions)
Tcs sp5 2, by Zeiss (1 mentions)
Eclipse te2000 u microscope, by Nikon (1 mentions)
Zen 2012, by Zeiss (1 mentions)
Upright axioimagerm2, by Zeiss (1 mentions)
Slide scannerz1, by Zeiss (1 mentions)
8 protocols using lsm780 inverted
1
Microscopic Cell Imaging Protocol
2
CRISPR Imaging of Human Telomeres
3
Immunofluorescence Staining of Cardiomyocytes
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Cells were fixed in 3.7% (v/v) paraformaldehyde for 10 min, then permeabilized and blocked in PBS with 4% (v/v) bovine growth serum (Thermo Scientific) and 0.2% (v/v) Triton X-100 for 1 h, and subsequently incubated with primary antibodies for AcH3 (Millipore), Actc1 (Sigma), sarcomeric MyHC (R&D), Nppa (Millipore), Ryr2 (Sigma), Pln (Abcam) in blocking buffer overnight on a rocker at 4 °C. Secondary antibody incubation (Alexa 647, Life Technologies) was performed in blocking buffer for 1 h at room temperature. The fixed PDMS membranes were then mounted on slides in Vectashield® mounting medium and analysed with confocal microcospy (Zeiss LSM-780 inverted) or conventional epifluoresence microscopy with a Zeiss Axio-Observer Z1 widefield fluorescent microscope.
High-throughput image analysis was performed with a custom made Matlab® script. Briefly, cell nuclei were detected using contrast enhancement methods, then segmented with an edge detection algorithm (Canny; Matlab®), following which elliptical fitting was performed to compute the aspect ratio (short axis/long axis) and the angle between the long axis and the groove direction.
High-throughput image analysis was performed with a custom made Matlab® script. Briefly, cell nuclei were detected using contrast enhancement methods, then segmented with an edge detection algorithm (Canny; Matlab®), following which elliptical fitting was performed to compute the aspect ratio (short axis/long axis) and the angle between the long axis and the groove direction.
4
Quantitative Analysis of ORF1 Expression
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Ten fields of view were taken on the Multiphoton Zeiss LSM780 inverted confocal microscope for each tumor and matched normal kidney. Images were deconvoluted with AutoQuant (AutoQuant) software using 10 iterations of three-dimensional deconvolution. The average fluorescence intensity of the ORF1 signal for each field of view was obtained using ImageJ software. The Z-stacks were projected using the sum slices projection type, and the mean gray value was recorded. The average fluorescence intensity of the ORF1 signal was normalized to the nuclei density. To obtain the nuclei volume using Imaris8 software (Bitplane), the surface of the blue channel was built using a baseline subtraction of 15,000. The ORF1 mean gray value was divided by the nuclei volume per field of view. To obtain the normalized fluorescence intensity, all images were then normalized to the average value of all 10 fields of view for the CMC87 sample.
5
Quantifying Fluorescence Intensity in Tissue Samples
6
Pollen Tube Germination Assay
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Transgenic pollen grains were germinated in liquid pollen tube germination medium (0.01% H3BO3, 5 mM CaCl2, 5 mM KCl, 1 mM MgSO4, 10% sucrose, pH 7.5) supplemented with 10 μM epibrassinolide. After incubation for 5 to 7 h at 23 °C, fluorescent images were captured using a Zeiss LSM780 inverted confocal microscope.
7
Biofilm Viability Analysis via CLSM
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To evaluate CLSM images, biofilms grown on dentin discs and treated with
different concentrations of BlueM® were washed and labeled with
Live/Dead BacLight Bacterial Viability Kit (L7012, Invitrogen Molecular Probes,
Eugene, OR, USA) containing SYTO9 (SY) and propidium iodide (PI), according to
the manufacturer’s instructions. The excitation/emission wavelengths were
488/500 nm for SYTO and 488/635 nm for PI. Fluorescence from the stained cells
was observed using a confocal laser scanning microscope (LSM 780 inverted,
Zeiss, Jena, Germany), and the images were acquired with ZEN 2012 software
(Zeiss) at a resolution of 1,024 by 1,024 pixels. Single focal plane images of
the biofilm were captured by the system with a 20x magnification lens. The area
selected for the biofilm analysis was randomly defined, but not so close to the
edges of the specimens.
different concentrations of BlueM® were washed and labeled with
Live/Dead BacLight Bacterial Viability Kit (L7012, Invitrogen Molecular Probes,
Eugene, OR, USA) containing SYTO9 (SY) and propidium iodide (PI), according to
the manufacturer’s instructions. The excitation/emission wavelengths were
488/500 nm for SYTO and 488/635 nm for PI. Fluorescence from the stained cells
was observed using a confocal laser scanning microscope (LSM 780 inverted,
Zeiss, Jena, Germany), and the images were acquired with ZEN 2012 software
(Zeiss) at a resolution of 1,024 by 1,024 pixels. Single focal plane images of
the biofilm were captured by the system with a 20x magnification lens. The area
selected for the biofilm analysis was randomly defined, but not so close to the
edges of the specimens.
8
Multimodal Imaging Techniques Protocol
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Imaging of in situ hybridization and X-gal staining was performed with Zeiss upright AxioImagerM2 through a 20X objective with tiling mode. HE staining was imaged with Zeiss upright AxioImagerM2 or Zeiss slide scannerZ1 through a 20X objective. For immunofluorescence and EdU labeling, images were acquired with Zeiss LSM780 inverted confocal microscope through a 20X objective. All images were processed with adobe Photoshop CS5.1.
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