Bright field images were captured on a Nikon SMZ 1500 microscope with a Spot Insight camera using Spot Basic software. Fluorescent and cytospin images were captured on a Nikon SMZ 1500 microscope with a Nikon Digital Sight camera or a Nikon Eclipse 80i microscope with a CRi Nuance Multispectral camera and a Nikon Digital Sight camera. Images were obtained using NIS Element AR 3.0 software or CRi Nuance 2.10.0 software. Composite images of different focal planes were compiled in Adobe Photoshop without altering the characteristics of the images. Confocal images were acquired on a Nikon Eclipse TE-2000E/C1 laser scanning confocal microscope using EZ-C1 3.80 software (Nikon) and analyzed with Fiji software (distributed by ImageJ, General Public License).
Smz1500 microscope
Manufactured by Nikon
Sourced in Japan
The Nikon SMZ1500 is a stereo microscope designed for a variety of applications. It features a zoom ratio of 15x and offers a wide field of view. The SMZ1500 is equipped with an Epi-Illumination system and provides high-quality optical performance.
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Lab products found in correlation
Hr plan apo 1x wd 54 objective, by Nikon (3 mentions)
Digital sight ds u1 camera, by Nikon (3 mentions)
Lsm 780 confocal microscope, by Zeiss (3 mentions)
Az100 microscope, by Nikon (2 mentions)
Tcs sp5 confocal microscope, by Leica (2 mentions)
Inverted a1r spectral confocal microscope, by Nikon (2 mentions)
Paraffin, by Merck Group (2 mentions)
Fast green, by Avantor (2 mentions)
Eclipse ts100 microscope, by Nikon (2 mentions)
Digital sight ds fi1 camera system, by Nikon (2 mentions)
Tms f microscope, by Motic (2 mentions)
Moticam 2300 3.0mp live resolution camera system, by Motic (2 mentions)
Ds u3, by Nikon (2 mentions)
T3 or t7 polymerase, by Thermo Fisher Scientific (2 mentions)
Model 308, by Aurora Scientific (2 mentions)
Model 400a, by Aurora Scientific (2 mentions)
Lsm 880, by Zeiss (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Lsm 510, by Zeiss (2 mentions)
Tricaine, by Merck Group (1 mentions)
92 protocols using smz1500 microscope
1
Microscopy Imaging Techniques Protocol
2
Analyzing Drosophila Wing Size and Fertility
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Lethal phase analysis was performed as described previously (Vasudevan et al., 2020 (link)). Right wings were severed from 1- to 4-day-old flies and imaged using a Nikon SMZ1500 microscope outfitted with a Nikon 8MP camera with NIS-Elements software. Wing size was measured using the regions of interest feature in ImageJ software.
Female fertility was quantified by placing five 1- to 4-day-old virgin females with five yw males (or Dj-GFP males for fertilization assays) in a vial containing standard medium enhanced with yeast to encourage egg laying. After allowing 1 day for acclimatization, the flies were moved to a new vial and the number of eggs laid in a 24-h period were counted and quantified. Eggs were imaged forFig. 6B by placing them on an apple juice plate and capturing them with a Nikon SMZ1500 microscope outfitted with 8MP Nikon camera controlled by NIS elements software. Ovaries from female flies in this experiment were dissected in ice-cold PBS and similarly imaged on apple juice plates for Fig. S4A .
Female fertility was quantified by placing five 1- to 4-day-old virgin females with five yw males (or Dj-GFP males for fertilization assays) in a vial containing standard medium enhanced with yeast to encourage egg laying. After allowing 1 day for acclimatization, the flies were moved to a new vial and the number of eggs laid in a 24-h period were counted and quantified. Eggs were imaged for
3
Fly Wing Size and Fertility Measurements
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Lethal phase analysis was performed as described previously (Vasudevan et al. 2020) .
Right wings were severed from 1-4 day old flies and imaged using a Nikon SMZ1500 microscope outfitted with a Nikon 8MP camera with NIS-Elements software. Wing size was measured using regions of interest (ROI) feature in ImageJ software.
was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint (which this version posted May 18, 2021. ; https://doi.org/10.1101/2021.05.18.444691 doi: bioRxiv preprint Female fertility was quantified by placing five 1-4 day old virgin females with five yw males in a vial containing standard media enhanced with yeast to encourage egg laying. After allowing a day for acclimatization, the flies were moved to a new vial and the number of eggs laid in a 24-hour period were counted and quantified. Eggs were imaged for Fig. 5b by placing them on an apple juice plate and captured with the Nikon SMZ1500 microscope outfitted with 8MP Nikon camera controlled by NIS elements software. Ovaries from female flies in this experiment were dissected in cold PBS and similarly imaged on apple juice plates for Fig. S3a.
Right wings were severed from 1-4 day old flies and imaged using a Nikon SMZ1500 microscope outfitted with a Nikon 8MP camera with NIS-Elements software. Wing size was measured using regions of interest (ROI) feature in ImageJ software.
was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint (which this version posted May 18, 2021. ; https://doi.org/10.1101/2021.05.18.444691 doi: bioRxiv preprint Female fertility was quantified by placing five 1-4 day old virgin females with five yw males in a vial containing standard media enhanced with yeast to encourage egg laying. After allowing a day for acclimatization, the flies were moved to a new vial and the number of eggs laid in a 24-hour period were counted and quantified. Eggs were imaged for Fig. 5b by placing them on an apple juice plate and captured with the Nikon SMZ1500 microscope outfitted with 8MP Nikon camera controlled by NIS elements software. Ovaries from female flies in this experiment were dissected in cold PBS and similarly imaged on apple juice plates for Fig. S3a.
4
Carmine Alum Staining of Mammary Glands
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Carmine alum staining of whole inguinal mammary glands was performed using standard protocols. Briefly, excised fourth and fifth mammary glands from the left and right side of adult approximately three-month-old rats were stretched and pressed onto a 2 × 3 in. glass slide and fixed 24 h in Carnoy’s fixative. Following rehydration to H2O, slides were stained in Carmine Alum stain 24 h and then serially dehydrated in ethanol. Finally, slides were cleared in Histo-Clear™ clearing reagent up to 72 h until translucent and coverslipped using Histomount mounting media. Images were taken on a Nikon SMZ1500 microscope.
5
Pharyngeal Contraction Rate Analysis
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Pumping rate was determined using a Sony camera attached to a Nikon SMZ1500 microscope that recorded 10 well fed day 1 adult animals per genotype. Pharyngeal contractions in 15 s time periods for 4 technical replicates were counted (by slowing video playback speed by 4×) for each animal using OpenShot and pumping rates per minute were calculated.
6
Zebrafish Thymic Fluorescence Assay
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For in vivo steroid sensitivity testing, 5-dpf larvae were sorted for thymic fluorescence, treated with dexamethasone (25 µg/ml, 0.25% ethanol) or ethanol vehicle alone (0.25%) in standard egg water, and imaged by fluorescent and light microscopy. For bright-field DIC images, a Zeiss Axio Imager.Z1 compound microscope equipped with an AxioCam HRc was used. For live imaging, zebrafish and larvae were anaesthetized using 0.016% tricaine (Sigma) and mounted in 4% methylcellulose (Sigma). A Nikon SMZ1500 microscope equipped with a Nikon digital sight DS-U1 camera was used for capturing both bright-field and fluorescent images from live zebrafish and larvae. For thymic fluorescence quantification, all animals in the same experiments were imaged at the same condition, and the acquired fluorescent images were quantified in ImageJ (NIH) by measuring the fluorescent-covered areas. To account for variation in fish size, the thymic fluorescent area was normalized against the bright-field area of the fish head. Overlays were created using ImageJ and Adobe Photoshop 7.0.1.
7
Root Twist and Angle Deviation
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Arabidopsis seedlings, germinated and grown as previously described, were treated with farnesene and/or taxol, both previously dissolved in ethanol (0.1% final volume) and added to the agar medium. The treatments consisted in 250 μM farnesene, 0.25 or 0.5 μM taxol or a combination of both compounds (0.25 μM taxol + 250 μM farnesene, and 0.5 μM taxol + 250 μM farnesene). Ethanol (0.1%) was used also in the control. Four replicates were performed for each treatment. Fourteen days after treatment, root meristems were analyzed with a Nikon SMZ 1500 microscope, and the presence of helical twist and the root angle deviation were evaluated and compared to the control by using the software Image Pro Plus (Media Cybernetics).
8
Jellyfish Bloom Monitoring Protocol
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The two specimens were collected using a 1.5×0.5 m Agassiz trawl from the Yellow Sea by the team investigating a project entitled “The key processes, mechanism and ecological consequences of jellyfish blooms in China coastal waters” in June 2012 (Qiu, 2014). They were fixed in ethanol and preserved in 75% ethanol. The specimens are deposited in the Marine Biological Museum of the Chinese Academy of Sciences (MBMCAS
9
Mating Confirmation and Body Size Analysis
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Mating set up was the same as described in Lifespan . We confirmed successful mating for all the worms by checking their progeny. With successful mating, about half of the progeny are male. For the only two sterile strains used in this study, ekl-1 and drh-3 homozygous mutants, we confirmed the successful mating empirically by observing the decrease in darkness of mated sterile worms under normal light, which indicates mating-induced fat loss. Images of live hermaphrodites on 60 mm plates were taken on Day 6/7 of adulthood with a Nikon SMZ1500 microscope. When RNAi was used, RNAi treatment always started from eggs for all the experiments. ImageJ was used to analyze the body size of the worms. The middle line of each worm was delineated using the segmented line tool and the total length was documented as the body length of the worm. A t-test was performed to compare the body size differences between groups of worms in the same day.
10
Prussian Blue Staining of Magnetically Labeled Cells
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24 h after magnetic labeling, the cells were collected by trypsinization, washed once with phosphate-buffered saline (PBS), and plated onto poly-lysine-coated glass slides using a cytospin (4 min, 1000 rpm). After drying the slides, the cells on the slides were fixed with ice-cold 4% paraformaldehyde for 30 minutes followed by two washing steps in PBS for 10 min each and incubation with a staining solution for 20 min. The staining solution was prepared immediately before use by mixing equal volumes of 20% hydrochloric acid and 10% potassium ferrocyanide solution. The slides were washed three times in distilled water and counterstained with nuclear fast red (Merck #15939) for 5 minutes. Thereafter, the slides were rinsed twice in distilled water, dehydrated in 95% ethanol and 2 changes of 100% ethanol, and mounted with Kaiser's Glycerine-Gelatine (#1.09242.0100, Merck). Images were acquired using a Nikon SMZ 1500 microscope.
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