Lsm800 confocal microscope

Manufactured by Zeiss

Sourced in Germany, United States, France, Japan, China, Italy, Australia, United Kingdom, Switzerland, Chile, Sweden

The ZEISS LSM 800 confocal microscope is a high-performance imaging system designed for advanced biological and materials research. It offers high-resolution, optical sectioning capabilities to visualize and analyze samples in three dimensions. The LSM 800 utilizes a laser-scanning technology to capture detailed images with excellent signal-to-noise ratio and contrast.

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1 273 protocols using lsm800 confocal microscope

Lysosomes and Phagocytic Hemocytes in Shrimp

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To label lysosome, the hemocytes were collected from 10 shrimps and incubated in staining solution (1 µl Lyso-Tracker Green (Beyotime, China), 133 µl Hoechst 33342 (Solarbio, China), 13.33 ml Insect-XPRESS™ (Lonza, Tampa, FL, USA)) for 1 h. After that, the hemocytes were washed with PBS for two times and observed with a LSM800 confocal microscope (ZEISS, Germany).
To label phagocytic hemocytes, overnight-cultured Vibrio parahaemolyticus (VP) was heat killed, labeled with 0.1 mg/ml FITC at 37°C for 1.5 h, washed and resuspended in PBS. The suspension was adjusted to 2 × 10⁸ particles/ml, and 100 μl suspension was injected into each shrimp. The hemocytes were collected from five injected shrimps, stained with Hoechst 33342 at 8 h postinjection and observed with a LSM800 confocal microscope (ZEISS, Germany).

Huang Z., Yang P, & Wang F. (2021). Shrimp Plasma CREG Is a Hemocyte Activation Factor. Frontiers in Immunology, 12, 707770.

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Quantification of Iba-1+ Immune Response

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Image acquisition of fluorescent staining images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.3 system) using the 10×, 20×, and 40× lenses. Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n). For analyses and bright field image acquisition of staining, Iba-1 and CSF1R, 8-bit grayscale TIFF images of the regions of interest were taken in a single sitting for whole protocols with a Qimaging camera (Qcapture program, version 2.9.10) attached to Nikon microscope (C-80) with the same gain/exposure settings for every image. To evaluate the level of Iba-1+ immune response in the regions of interest, the images were imported into ImageJ (1.37) and the percentage of area occupied by the staining was measured using the threshold parameter. Cell count was assesed manually using ImageJ (1.37). Analysis was performed in double blinded to avoid bias of analysis. Fluorescent staining of images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.6 system). Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n).

Pons V., Lévesque P., Plante M.M, & Rivest S. (2021). Conditional genetic deletion of CSF1 receptor in microglia ameliorates the physiopathology of Alzheimer’s disease. Alzheimer's Research & Therapy, 13, 8.

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Subcellular Localization and Protein-Protein Interactions of Soybean IQM Genes

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The protoplasmic transformation for both experiments was performed as previously described (Lv et al., 2019 (link)). For subcellular localization, the full-length CDS of 15 IQM genes in soybean Williams82 was cloned into the pGreenII-35S-GFP plant expression vector, respectively. Thereafter, pGreenII-35S-GFP-GmIQMs plasmids were transformed into protoplasts isolated from the leaves of 4-week-old Arabidopsis wild-type Col-0. After 16 h of incubation at 22°C, the GFP signal was observed using a Zeiss LSM800 confocal microscope at 488 nm absorption and 507 nm emission wavelengths. The pGreenII-35S-GFP empty vector was used as a negative control.
For the BiFC assays, the CDSs of GmIQM1d/GmIQM2c and GmCaM were cloned into binary pSAT1-nEYFP and pSAT1-cEYFP vectors, respectively. Thereafter, the pSAT1-nEYFP-GmIQM1d/GmIQM2c and pSAT1-cEYFP-GmCaM plasmids were transformed into protoplasts isolated from the leaves of 4-week-old Col-0. After 16 h of incubation at 22°C, the EYFP signal was observed using a Zeiss LSM800 confocal microscope at 488 nm absorption and 530 nm emission wavelengths. Co-transformation with nEYFP empty vector and cEYFP-GmCaM served as negative controls.

Lv T., Liu Q., Xiao H., Fan T., Zhou Y., Wang J, & Tian C.E. (2023). Genome-wide identification and analysis of the IQM gene family in soybean. Frontiers in Plant Science, 13, 1093589.

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Quantitative Microscopy of Immune Markers

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Image acquisition of Fluorescent staining images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.3 system) using the 10×, 20×, and 40 × lenses. Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n). For analyses and bright eld image acquisition of staining, Iba-1 and CSF1R, 8-bit grayscale TIFF images of the regions of interest were taken in a single sitting for whole protocols with a Qimaging camera (Qcapture program, version 2.9.10) attached to Nikon microscope (C-80) with the same gain/exposure settings for every image. To evaluate the level of Iba-1 + immune response in the regions of interest, the images were imported into ImageJ (1.37) and the percentage of area occupied by the staining was measured using the threshold parameter. Cell count was assed manually using ImageJ (1.37). Analysis was performed in double blinded to avoid bias of analysis. Fluorescent staining of images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.6 system). Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n).

Pons V., Lévesque P., Plande M., & Rivest S. (2020). Conditional genetic deletion of CSF1 receptor in microglia ameliorates the physiopathology of Alzheimer’s disease.

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High-resolution Confocal Imaging Workflow

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Laser confocal experiments were acquired using a Zeiss LSM 800 confocal microscope equipped with a 63×1.4 numerical aperture oil objective. Airyscan microscopy was performed using a Zeiss LSM 800 confocal microscope, equipped with Plan-Apochromat 63×/1.4 numerical aperture oil objective and pixel size of 8.7 nm. Images were subjected to post-acquisition Airyscan processing. Image acquisition and processing were performed with Zen Blue software and co-localization analysis and image presentation was performed using Image J FIJI software or Photoshop (Adobe).

Lin Z., Ni L., Teng C., zhang z., Lu X., Xie C., Wu L., Zhou Y., Tian N., Wu Y., Sun L., Pan Z., Wang X., Lin Z., & Zhang X. (2021). Nrf2 mediated ER-phagy protects against oxidative damage in intervertebral disc degeneration.

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Quantifying Iba-1+ Microglial Response

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Image acquisition of Fluorescent staining images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.3 system) using the 10×, 20×, and 40× lenses. Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n). For analyses and bright eld image acquisition of staining, Iba-1 and CSF1R, 8-bit grayscale TIFF images of the regions of interest were taken in a single sitting for whole protocols with a Qimaging camera (Qcapture program, version 2.9.10) attached to Nikon microscope (C-80) with the same gain/exposure settings for every image. To evaluate the level of Iba-1+ immune response in the regions of interest, the images were imported into ImageJ (1.37) and the percentage of area occupied by the staining was measured using the threshold parameter. Cell count was assed manually using ImageJ (1.37). Analysis was performed in double blinded to avoid bias of analysis. Fluorescent staining of images was performed using a Zeiss LSM800 confocal microscope supported by the Zen software (2.6 system). Confocal images were then processed using Fiji (ImageJ Version 2.0.0-rc-43/1.51n).

Pons V., Lévesque P., Plande M., & Rivest S. (2020). Conditional Genetic Deletion of CSF1 Receptor in Microglia Ameliorates the Physiopathology of Alzheimer’s Disease.

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Confocal Imaging of Embryos

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Embryos were imaged with a Zeiss LSM800 confocal microscope; cells were imaged by confocal or with an inverted Zeiss AxioObserver. For time-lapse imaging, dechorionated St12 embryos were mounted in halocarbon oil and scanned at 6 min intervals. For single-frame live imaging, embryos were dechorionated, mounted in PBT, and directly scanned. Control and mutant embryos were prepared and imaged in parallel where possible, and imaging parameters were maintained between genotypes. The fluorescent intensity and cell morphology measurements were made with ImageJ software.

Yang S., McAdow J., Du Y., Trigg J., Taghert P.H, & Johnson A.N. (2022). Spatiotemporal expression of regulatory kinases directs the transition from mitosis to cellular morphogenesis in Drosophila. Nature Communications, 13, 772.

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Fluorescence Microscopy for Diverse Assays

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Samples for immunocytochemistry, PLA, TUNEL staining, and GFP-based reporter analyses were subjected to fluorescence microscopy. Images for immunocytochemistry and PLA (in neurons) and TUNEL-stained cell death analysis were acquired using a 40× oil objective on an Axio Observer.Z1 inverted microscope (Zeiss) equipped with an AxioCam MRm Rev 3. camera. PLA puncta within the region of interest (ROI) were counted manually and normalized to ROI area. For image generation for PLA in brain sections and GLI activity-dependent expression of GFP in neurons, a LSM800 confocal microscope (Zeiss) with a 40x or 63x oil objective was employed. For tissue PLA, images were acquired in Z stacks and tiles to generate comprehensive images of greater depth and larger field of view. PLA puncta within the ROI were counted and normalized by ROI area using Fiji/ImageJ software.

Chung K.M., Kim H., Roque C.G., McCurdy E.P., Nguyen T.T., Siegelin M.D., Hwang J.Y, & Hengst U. (2022). A systemic cell stress signal confers neuronal resilience toward oxidative stress in a Hedgehog-dependent manner. Cell reports, 41(3), 111488.

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Measuring Mitochondrial Membrane Potential

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To measure the mitochondrial membrane potential, adipocytes were stained with cell growth medium containing 100 nM tetramethylrhodamine, methyl ester (TMRM, I34361, Invitrogen). Cells were then incubated for 30 min at 37°C. The images were captured with a ZEISS LSM800 confocal microscope.

Tan X., Zhu T., Zhang L., Fu L., Hu Y., Li H., Li C., Zhang J., Liang B, & Liu J. (2022). miR-669a-5p promotes adipogenic differentiation and induces browning in preadipocytes. Adipocyte, 11(1), 120-132.

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Immunostaining of Serotonin in Fly Brains

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For all immunostainings, adult female flies 5–7 days after eclosion fed with 5-HTP or normal food were anesthetized, and their brains were dissected in ice-cold PBS and fixed in 4% paraformaldehyde in PBS for 30 min at room temperature, followed by 4 rinses in PAT3 (0.5% Triton X-100, 0.5% bovine serum albumin in PBS) for 10 min at room temperature. Samples were transferred to 5% normal donkey serum (NDS) or normal goat serum (NGS) in PAT3 for 1 h of blocking at room temperature and incubated with primary antibodies (diluted in 5% NDS or NGS) at room temperature for 4 h and then at 4 °C overnight. After washing samples 4 times for 10 min with PAT3 at room temperature and incubating samples with secondary antibodies in 5% NDS or NGS at room temperature for 4 h and 4 °C overnight, we mounted the samples with FocusClear^™ (Cat# FC-10100, CelExplorer Labs, Taiwan, China) and imaged them on a Zeiss LSM800 confocal microscope. The following antibodies were used: rabbit anti-5-HT (1:1000; Cat# 20080, RRID: AB_572263, ImmunoStar, Hudson, WI, USA) and mouse anti-nc82 (1:20; Cat# 2314866, RRID: AB_2314866, DSHB, USA). Secondary antibodies were diluted at 1:500 and were as follows: goat anti-rabbit Alexa Fluor 488 (Cat# A11008, RRID: AB_143165, Thermo Fisher Scientific, Foster City, CA, USA) and donkey anti-mouse Alexa Fluor 568 (Cat# A10037, RRID: AB_2534013, Thermo Fisher Scientific).

Cao H., Tang J., Liu Q., Huang J, & Xu R. (2022). Autism-like behaviors regulated by the serotonin receptor 5-HT2B in the dorsal fan-shaped body neurons of Drosophila melanogaster. European Journal of Medical Research, 27, 203.

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