Leica system

Manufactured by Leica camera

The Leica System is a comprehensive suite of high-precision camera equipment and accessories designed for professional and advanced amateur photographers. The core function of the Leica System is to capture and record images with exceptional quality, detail, and clarity. The system includes a range of interchangeable lenses, cameras, and other accessories that work seamlessly together to provide a versatile and reliable imaging solution.

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Lab products found in correlation

Rhodamine dextran, by Thermo Fisher Scientific (1 mentions) Qtracker 655, by Thermo Fisher Scientific (1 mentions) Qtracker 655 vascular label, by Thermo Fisher Scientific (1 mentions) Rhodamine dextran, by Merck Group (1 mentions) Dmem medium, by Nacalai Tesque (1 mentions) Fugene hd, by Promega (1 mentions)

5 protocols using leica system

Time-lapse Microscopy Image Processing

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The software implementation of our method was run in GNU Octave, version 3.2.4, compatible with MATLAB. The program tiffread.m[30] (link) was used for converting the TIFF images obtained from the time-lapse microscopy into real matrices.
A detailed description of the software is provided in Documents S1-S3.Image files from the Leica system (.lif) have been handled with ImageJ that has also been used to produce “Sum slice” projections [31] .

Zambrano S., Bianchi M.E, & Agresti A. (2014). High-Throughput Analysis of NF-κB Dynamics in Single Cells Reveals Basal Nuclear Localization of NF-κB and Spontaneous Activation of Oscillations. PLoS ONE, 9(3), e90104.

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Intravital Multiphoton Imaging of Bone Marrow

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Mice were prepared for intravital TPLSM as previously described^{4 (link)}. TPLSM was performed with a Leica system as described above. EGFP⁺ cells of 7-12 week old female LysM-EGFP mice and tdTomato⁺ cells of Catchup^IVM-red mice were excited at 960nm, at which bone tissue additionally emits a second-harmonic generation (SHG) signal at 480nm. Fluorescent cells were detected with specific filters at 525/50nm (EGFP) or 585/50nm (tdTomato) and SHG was detected via a 460/50nm filter.
Blood flow was visualized by injecting 1.5mg/ml Rhodamine Dextran (Sigma-Aldrich, Cat# R9379-100MG) or 1μM Qtracker™ 655 Vascular Labels (Thermo Fisher, Cat# Q21021MP) in a total volume of 100μl PBS i.v.. Fluorescence was excited at 960nm and detected with a 585/40nm (Rhodamine Dextran) or a 650/50nm (Qtracker 655) filter. Imaging was performed as well in resonant as in non-resonant detection mode. Scan speed was adjusted individually for different vessel types from 600 Hz to 12 KHz.
Neutrophils were activated by injecting 100μg/kg body weight hG-CSF (Neupogen®, Amgen GmbH) i.v. in a total volume of 100μl PBS. The raw data were reconstructed and analyzed using Imaris software (Bitplane) and ImageJ.
Further information on software versions used for data collection and processing are listed in the reporting summary document and Supplementary table 4.

Grüneboom A., Hawwari I., Weidner D., Culemann S., Müller S., Henneberg S., Brenzel A., Merz S., Bornemann L., Zec K., Wuelling M., Kling L., Hasenberg M., Voortmann S., Lang S., Baum W., Ohs A., Kraff O., Quick H.H., Jäger M., Landgraeber S., Dudda M., Danuser R., Stein J.V., Rohde M., Gelse K., Garbe A.I., Adamczyk A., Westendorf A.M., Hoffmann D., Christiansen S., Engel D.R., Vortkamp A., Krönke G., Herrmann M., Kamradt T., Schett G., Hasenberg A, & Gunzer M. (2019). A network of trans-cortical capillaries as mainstay for blood circulation in long bones. Nature metabolism, 1(2), 236-250.

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Quantitative Image Analysis of Fibrosis

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Quantification of fibrosis, immunohistochemistry, and immunofluorescence analysis was performed using ImageJ software (NIH Image, Bethesda, MD, USA). Five randomly chosen high-resolution images per mouse (original magnification ×100) captured using the Leica system were used for quantification. Two independent investigators, blinded to the experimental conditions, performed this process.

Zheng H., Zhang Y., He J., Yang Z., Zhang R., Li L., Luo Z., Ye Y, & Sun Q. (2021). Hydroxychloroquine Inhibits Macrophage Activation and Attenuates Renal Fibrosis After Ischemia-Reperfusion Injury. Frontiers in Immunology, 12, 645100.

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Amniotic Fluid Karyotyping Protocol

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Approximately 20 ml of amniotic fluid was centrifuged at 1,500 rpm for 10 min at 4°C to separate amniotic cells. The cells were cultured, harvested and mounted on glass slides for the chromosome G-banded analysis by the Leica system. On each slide, 20 stained metaphases were randomly selected and examined, and 6 karyograms were created for the chromosome analysis. If a suspicious chromosomal abnormality or a chromosomal polymorphism was found, the count number of metaphases was then increased to a number between 50 and 100 for a more reliable result.

Sun S., Zhang Z., Zhao J, & Lan X. (2022). Extended application of BACs-on-Beads technique in prenatal diagnosis. Archives of Medical Science : AMS, 19(1), 250-257.

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RBM20 Mutant Protein Impacts

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HeLa and HEK293T cells were cultured in DMEM Medium (Nacalai) supplemented with 10% fetal bovine serum at 37 °C with 5% CO₂. Plasmid DNAs were transfected by using FuGENE HD (Promega) according to the manufacturer’s instruction. The Ttn reporter minigene and the expression vectors for FLAG-tagged wild-type or mutant RBM20 proteins were co-transfected in a 1:4 mixture. Fluorescence images of fluorescent proteins were acquired 24–36 hours after transfection by using Leica system, and then, the cells were harvested for total RNA preparation and RT-PCR analysis

Murayama R., Kimura-Asami M., Togo-Ohno M., Yamasaki-Kato Y., Naruse T.K., Yamamoto T., Hayashi T., Ai T., Spoonamore K.G., Kovacs R.J., Vatta M., Iizuka M., Saito M., Wani S., Hiraoka Y., Kimura A, & Kuroyanagi H. (2018). Phosphorylation of the RSRSP stretch is critical for splicing regulation by RNA-Binding Motif Protein 20 (RBM20) through nuclear localization. Scientific Reports, 8, 8970.

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