Immunofluorescence images were captured using a 100 X (1.4 N.A.) oil-immersion objective attached on a Leica SP 5 laser scanning inverted confocal microscope. The same image acquisition settings were used for both control and the mutant EB1-treated conditions. The images were analyzed using Leica Application Suite Advanced Fluorescence Lite software. To measure the wild type and the mutant EB1-GFP intensity on the rhodamine-labeled microtubules, region of interest (ROI) lines were drawn across the length of the individual microtubules and the GFP intensity was measured after subtracting the background of same area for each. A representative image showing the ROI is provided in the Supplementary Fig. S11 . The resultant mean intensity per pixel of wild type EB1-GFP and the mutant EB1-GFP bound to the microtubules were plotted. About 30 microtubules were analyzed for each case to obtain the statistical significance.
Application suite advanced fluorescence lite software
Manufactured by Leica
Sourced in Germany
The Application Suite Advanced Fluorescence Lite software is a core imaging and analysis tool for Leica's microscopy equipment. It provides essential functionalities for fluorescence microscopy, enabling users to capture, process, and analyze fluorescence-based images and data.
Automatically generated - may contain errors
Lab products found in correlation
Sp5 inverted confocal laser scanning microscope, by Leica (1 mentions)
Axiocam mr, by Zeiss (1 mentions)
Axioskop 2 microscope, by Zeiss (1 mentions)
X63 oil immersion objective, by Zeiss (1 mentions)
Tcs sp5 2, by Leica (1 mentions)
Sp8 x inverted confocal microscope, by Leica (1 mentions)
Tcs sp8 x confocal microscope, by Leica (1 mentions)
Cellmask orange plasma membrane stain, by Thermo Fisher Scientific (1 mentions)
Vectashield, by Vector Laboratories (1 mentions)
5 protocols using application suite advanced fluorescence lite software
1
Quantifying Wild Type and Mutant EB1 Binding
2
Microscopic Imaging of Nematodes
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DIC (Nomarski) and fluorescent imaging was carried out using a Zeiss AxioSKOP2 microscope with a Zeiss AxioCamMR digital camera. Photomicrographs were taken using a x63 oil immersion objective (Zeiss) and Axiovision software (Release 4.5). Confocal images were taken on a Leica TCS SP5II using Leica Application Suite Advanced Fluorescence Lite software (Release 2.2.1). In both cases, animals were mounted on agarose pads (2% agarose, 0.5% 1-phenoxy-2-propanol in M9) in 0.2% 1-phenoxy-2-propanol.
3
Visualizing Protein Localization in Plant Cells
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Agroinfiltrated leaf sections were imaged at room temperature using a Leica SP8 X inverted confocal microscope with an argon laser (Leica, Wetzlar, Germany). DAPI was excited at 340–488 nm, and the emitted light was captured at 505–555 nm. GFP was excited at 488 nm, and the emitted light was captured at 505–555 nm. YFP was excited at 514 nm, and the emitted light was captured at 530–590 nm. Images were captured digitally and processed using the Leica Application Suite Advanced Fluorescence Lite software (LAS AF; version 2.6.3; build 8173).
4
Confocal Imaging of Agroinfiltrated Leaves
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Leica SP8 X inverted confocal microscope with an argon laser (Leica, Wetzlar, Germany) was used to image the agroinfiltrated leaf sections at room temperature. YFP was excited at 514 nm, and the emission was captured at 530–590 nm. The RFP was excited at 561 nm, and the emission was captured at 588–648 nm. Images were captured digitally and processed using Leica Application Suite Advanced Fluorescence Lite software (Leica Microsystems). ImageJ (http://rsbweb.nih.gov/ij/ ) was used to quantify the fluorescence intensity.
5
Visualizing Antigen-Presenting Cells in Mice
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The transgenic mice C57BL/6 that express the green fluorescent (GFP) under the major histocompatibility complex class II molecule (MHC-II) promoter were i.d. injected in the ears with PBS, iC (1 × 106 iC) stained with the CellMask Orange Plasma Membrane Stain (Thermo Fisher Scientific, United States), or LC (1 × 106 LC) stained with CellMask Orange Plasma Membrane Stain. Six hours later the ears were removed and treated with 0.5 M EDTA for 2 h and then were washed with PBS for 20 min. The epidermal layer was then separated from the dermal layer, washed, and acetone-fixed for 20 min at -20°C. Afterward, the epidermal sheets were mounted with VectaShield (Vector Laboratories, Burlingame, CA, United States) and sealed. The images were obtained with a Leica TCS SP8x Confocal Microscope (Wetzlar, Germany) and analyzed with Leica Application Suite Advanced Fluorescence Lite software (Leica Microsystems, Mannheim, Germany).
For the staining, 1 × 106 conidia of S. schenckii were incubated with 1000 μL of CellMask Orange 1× for 30 min at 37°C, after that the conidia were washed twice with PBS (PBS).
For the staining, 1 × 106 conidia of S. schenckii were incubated with 1000 μL of CellMask Orange 1× for 30 min at 37°C, after that the conidia were washed twice with PBS (PBS).
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