L4 animals treated with 2.8 μM of purified tCPR-4 proteins in liquid culture for 48 hours were transferred to NGM plates and allowed to recover for 1 hour at 20°C. They were then dissected to expose their gonads following the protocol described previously24 (link). Dissected gonads were fixed and stained with DAPI34 (link). The number of germ nuclei and the number of metaphase nuclei in the mitotic zone of each gonad were scored using a Zeiss Nomarski microscope with a DAPI filter24 (link).
Nomarski microscope
Manufactured by Zeiss
The Nomarski microscope is an optical microscope that utilizes a specialized illumination technique called differential interference contrast (DIC) to enhance the visibility of unstained, transparent specimens. It allows for the observation of finer details and surface topography in samples by creating a three-dimensional, shadow-like effect. The Nomarski microscope is commonly used in various scientific and research applications that require high-contrast imaging of delicate samples.
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Lab products found in correlation
4 protocols using nomarski microscope
1
Quantification of Germ Cell Nuclei
2
Whole-Mount Root Clearing and Flower Imaging
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For whole-mount observation, the roots were cleared and mounted as described [23] (link). Briefly, roots were fixed in 0.24N HCl with 20% methanol and incubated at 57°C for 15 minutes. After that, the solution was replaced to 7% NaOH, 7% hydroxylamine-HCl in 60% ethanol at room temperature for 15 minutes. Roots were then rehydrated through conventional ethanol series (40%, 20% and 10%) with 5 minutes each step, and cleared in 5% ethanol, 25% glycerol. Roots were soaked in 50% glycerol to observation by a Nomarski microscope (Zeiss).
To photograph flowers, flowers of appropriate stage were dissected under an Olympus AX-70 microscope and photographed. Image pro software was used to measure the length of pistil and stamens.
To photograph flowers, flowers of appropriate stage were dissected under an Olympus AX-70 microscope and photographed. Image pro software was used to measure the length of pistil and stamens.
3
Quantification of Germ Cell Nuclei
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L4 animals treated with 2.8 μM of purified tCPR-4 proteins in liquid culture for 48 hours were transferred to NGM plates and allowed to recover for 1 hour at 20°C. They were then dissected to expose their gonads following the protocol described previously24 (link). Dissected gonads were fixed and stained with DAPI34 (link). The number of germ nuclei and the number of metaphase nuclei in the mitotic zone of each gonad were scored using a Zeiss Nomarski microscope with a DAPI filter24 (link).
4
Imaging-based Stress Reporter Analysis
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For imaging analysis, animals were immobilized with 5% sodium azide (UN3287 ICCA) at the indicated time point. For HSP-4p::GFP, NLP-29p::GFP and GCS-1p::GFP stress reporter inductions, images were acquired using an Axiozoom microscope (Zeiss). For DHS-3::GFP LD visualization, images were acquired using a Nomarski microscope (Zeiss running Zen v.2.5) with ApoTome. All fluorescence quantifications were processed using Fiji (ImageJ v.1.54f). For quantification, the animal boundary was defined using bright-field imaging and the mean pixel intensity was calculated using a fluorescent image taken with the same field of view. Quantification was presented as FC to WT animals at 0 h for a given treatment. For confocal microscopy of Grx1-GFP2 strains, images were acquired using a Zeiss LSM900 equipped with Airyscan 2, GaAsP PMT detectors and 405/488/561/640-nm lasers running Zen v.3.4. Specifically, excitation by a 405-nm laser and a 488-nm laser was performed in tandem to determine the relative fractions of oxidized and reduced Grx1-roGFP2, respectively. Emissions were detected by a GaAsP PMT with no filters.
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