Fission planes were revealed by compression between a plastic tissue culture dish and a glass coverslip (See Video S3 ). Animals were inverted with their ventral side up, compressed using four fingertips, then imaged. To ensure that all compression/fission planes were revealed for every animal, images were acquired sequentially using a Leica M205 microscope as each fission plane was revealed by mechanical compression. Position of fission planes and distance between fission planes was quantified using Fiji (https://fiji.sc /). Video depicting compression assay was captured with an iPhone 6 (Apple).
M205 microscope
Manufactured by Leica
Sourced in Germany
The Leica M205 is a high-performance stereomicroscope designed for a wide range of applications. It features a zoom range of 2x to 20.5x, providing versatile magnification capabilities. The M205 is equipped with LED illumination, ensuring consistent and uniform lighting for clear and detailed observations.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse ti, by Yokogawa (2 mentions)
W1 spinning disk, by Yokogawa (2 mentions)
Zen black software, by Nikon (2 mentions)
Dfc450, by Leica (1 mentions)
Creative suite, by Adobe (1 mentions)
Dfc290, by Leica (1 mentions)
Tricaine, by Merck Group (1 mentions)
Orca flash 4 v3 camera, by Nikon (1 mentions)
Ti2 inverted microscope, by Yokogawa (1 mentions)
Mc190 hd, by Leica (1 mentions)
17 protocols using m205 microscope
1
Revealing Fission Planes via Compression
2
Multimodal Imaging for Comprehensive Gene Expression Analysis
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Colorimetric whole-mount in situ hybridization samples and live worm or fragment images were acquired using a Leica M205 microscope using the Leica Application Suite (LASX). Following image acquisition, non-tissue background was subtracted, contract and image intensity adjusted, and the edited image was converted to grayscale for data presentation. No quantifications were performed on contrast or intensity adjusted images and all raw, original data is available in the original data repository (http://www.stowers.org/research/publications/libpb-1513 ). Confocal images of fluorescent in situ hybridization samples were acquired using an LSM-700 inverted confocal microscope with Zeiss Zen Black Software (v8.1) or via high throughput imaging on a Nikon Eclipse Ti with a Yokogawa W1 spinning disk and robotic plate loader. With automated image capture, whole slides were imaged at 4X and objects of interest were automatically detected, re-imaged at 10X, then batch stitched, quantified and aligned using Fiji macros (https://github.com/jouyun/smc-macros ). Stitching of tiles for whole worm images was performed with Fiji plugins (grid/collection stitching) using custom macros for batch processing. Maximum intensity projections of the stitched z-stacks were generated to visualize gene expression across the entire animal, then rotated, and cropped for data presentation.
3
Microscopic Imaging of Worm Regeneration
4
Imaging RNAi-Treated Planarians
5
Revealing Fission Planes via Compression
6
Zebrafish Xenograft Assay for Cancer Cell Dissemination
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Adult zebrafish were maintained in a zebrafish facility with a 14:10 day:night cycle and handled in compliance with an approved institutional protocol. Control NMuMG cells and cells with down-regulated ShcA expression were grown to confluence, washed twice with PBS, trypsinized and labeled with CM-DiI by immersion for 5 min at 37°C, and then transferred to ice for 15 min, as instructed by the manufacturer. Cells were then washed three times with PBS, suspended in PBS, and transferred into a borosilicate needle for injection into anesthetized dechorionated embryos, held on agarose-lined plates. 100–150 DiI-labeled cells were injected in the mid yolk sac region of the zebrafish embryos. After injection, embryos were sorted for fluorescence, and pictures were taken. Subsequently, the xenografted embryos were held at 31–32°C for 3 d prior to imaging of cell dissemination. Larvae were anesthetized with 0.003% tricaine (Sigma), and pictures were taken using a Leica M205 microscope. For each condition, the data shown are from at least two experiments with at least fifty embryos per group. Images were analyzed and modified for brightness and contrast using Adobe Photoshop CS5 software.
7
Teratogenic Effects of DEB on Zebrafish
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The embryos obtained from indicated breeding crosses were treated with DEB (Sigma Aldrich) at indicated concentrations in egg water with methylene blue between 4 and 72 hpf. The embryos obtained from heterozygous mutant crosses were separated at the end of treatment into three groups based on the severity of the observed morphological changes (normal, moderate and severe) and genotyped using fPCR as described earlier. Representative images of DEB treated embryos obtained from homozygous mutant inbreeding or outbreeding were taken using LAS X Imaging software on a Leica M205 microscope with a DFC7000 color camera.
8
Confocal Imaging of Zebrafish Intracranial Lymphatics
9
Microscopic Imaging of Worm Regeneration
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Images of live worms and regenerating fission fragments were acquired using a Leica M205 microscope. Confocal images were acquired on an LSM-700-Vis and stitching was performed in Fiji using built-in grid collection plugins.
10
Measurement of Fly Strain Pigmentation
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Strain pigmentation phenotypes were measured by collecting flies without CO2 anesthetization on the day of eclosion and aging 5 days at 25°C to normalize cuticular tanning. We then anesthetized aged flies, mounted the adult abdomens to double-sided sticky tape on slides, and imaged the abdomen using standard settings on a Leica M205 microscope. Images were quantified using the ImageJ program [54 (link)] to measure the area of the tergite that was pigmented, divided by the total area of the tergite, yielding a percent pigmentation score. The relationship between strain pigmentation and latitude, longitude, and altitude was analyzed by nominal logistic regression.
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