The morphology of the isolates was assessed according to Landolt 1986 [1 ] as well as Les et al. [11 (link)]. An M205 FCA fluorescence stereo microscope (Leica, Wetzlar, Germany) and Axio Imager 2 light microscope (Zeiss, Jena, Germany) was used.
M205 fca fluorescence stereo microscope
Manufactured by Leica
Sourced in Germany
The Leica M205 FCA is a fluorescence stereo microscope designed for high-resolution imaging. It features a large zoom range, high-performance optics, and a state-of-the-art camera system for capturing detailed images of fluorescently labeled samples.
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Lab products found in correlation
6 protocols using m205 fca fluorescence stereo microscope
1
Microscopic Analysis of Microalgae Isolates
2
Hydrogen Peroxide Visualization with PO1
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Hydrogen peroxide was visualized with PO1. PO1 (Tocris) was dissolved in DMSO to make a 500 μM stock and was further diluted in water to make a 40 μM working solution. Seven days-old seedlings were incubated in PO1 for 5 min in the dark and were then rinsed with water and mounted in water for imaging. Seedlings were imaged on a Leica M205 FCA fluorescence stereo microscope using a 545-620nm filter.
3
Arabidopsis thaliana Genetic Mutant Characterization
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All Arabidopsis thaliana plants used in this study were in the Col‐0 ecotype background. The hst‐1, hst‐3, ago1‐18, ago1‐27, hyl1‐2, se‐1, dcl2‐1, dcl3‐1, dcl4‐2, hen1‐6, and shr‐2 mutant lines and pSUC2::GFP, pSUC2::tmGFP9, and SUC‐SUL transgenic lines were described previously (Fukaki et al, 1998 (link); Telfer & Poethig, 1998 (link); Imlau et al, 1999 (link); Prigge & Wagner, 2001 (link); Morel et al, 2002 (link); Bollman et al, 2003 (link); Himber et al, 2003 (link); Vazquez et al, 2004a (link); Xie et al, 2004 (link); Li et al, 2005 ; Sorin et al, 2005 (link); Stadler et al, 2005 (link); Xie et al, 2005 (link)). Surface sterilized seeds were sown on ½MS medium containing MES buffer and vitamins (Duchefa Biochemie) and solidified with 0.8% microagar. Plants were cultivated in vitro at 21°C in 12‐h light/12‐h dark conditions for two weeks before transplanting in soil. Further growing was done at 21°C in 16‐h light/8‐h dark conditions. Light intensity was 120 µE.m−2.s−1 in every condition. Plant phenotype pictures were taken on 4‐week‐old plants, with the exception of in vitro cultured plants, for which phenotyping pictures of 2‐week‐old plants were taken using a M205 FCA fluorescence stereo microscope (Leica).
4
Quantifying Anti-pH3-PS10 Immunolabeling
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For imaging of anti-pH3-PS10 immunolabeled samples, z-stacks of mounted animals (tail region, dorsal side up) were collected on a Leica M205 FCA Fluorescence stereo microscope. The magnification was set at 8.04X, DAPI (ET DAPI) and pH3-PS10 (ET mCHER) channels we used. The same exposures were maintained across samples. After image acquisition parallax correction was applied in the LAS × software (version 3.4.1.17822). Animal area and number of PS10+ cells were quantified in FIJI running ImageJ 1.53t40 . Briefly, a z-projection was created from the parallax files by selecting the max intensity for projection type, and the channels were split. “Auto Local Threshold” with method Otsu and default settings was used on the DAPI image to identify the animal area, which was outlined using the wand tool, manually cropped to the post-pharyngeal region if necessary, then measured. pH3-PS10+ cells were counted using the “Find Maxima” function within the outline generated with the DAPI channel. The area and pH3-PS10 count results were exported to excel where the number of mitosis per mm2 were calculated.
5
Zebrafish Embryo Anesthesia and Imaging
6
Pollen Morphometric Analysis Protocol
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100 to 600 uL freshly shed pollen was collected in tassel bags before being added to individual microcentrifuge tubes containing 800 uL EAA fixative solution (3:1 ethanol to acetic acid). Pollen samples were inverted 3x before being parafilmed and stored at 20 C. Samples were imaged using a LEICA M205 FCA Fluorescence stereo microscope and the Leica Application Suite X (V 3.7.5.24914). Resulting images were imported into Fiji (ImageJ) (V 2.0.0) and subjected to the following processing pipeline: ‘Image -> Enhance Contrast’ (0.3%), ‘Adjust - >Threshold Image’ (Auto -> Apply), ‘Process -> Binary -> Make Binary’, ’Process -> Binary -> Watershed’, ‘Analyze -> Analyze Particles (Size (micron^2): 2000–14000, Circularity: 0.75–1.00, Show:Nothing, Display Results, Summarize, Exclude on edges, Include holes) -> Okay’. Particle measurements copied into a .csv file and loaded into R (V. 4.2.3) to produce plots with ggplot2 (V 3.4.3) and ggridges (0.5.4) packages.
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