To evaluate EB growth dynamics and patterning, the EBs (24 EBs/per stage; three experiments; N = 288 for each cell line) cultivated in 12 or 96-well plates with a low-attachment surface (Greiner Bio-One International GmbH and Nunclon Sphera; Nuclon) and imaged at the studied stages. The EB images were captured using Olympus CK40 inverted microscope equipped with a Camedia C-4040 camera (Olympus). EB diameters were measured in 2-8 directions for average calculation using ImageJ/Fiji software (https://imagej.nih.gov/ij/ ) with prior set up absolute scale calibration (see Supplementary Methods ). EB volumes, volumetric pattern sizes and their ratios were calculated using Excel.
Camedia c 4040 camera
Manufactured by Olympus
The Camedia C-4040 is a digital camera manufactured by Olympus. It features a 4.1-megapixel CCD sensor and a 3x optical zoom lens. The camera supports various image file formats, including JPEG and TIFF.
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2 protocols using camedia c 4040 camera
1
Evaluating Embryoid Body Growth Dynamics
2
Histological Tissue Processing and Staining
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Tissue samples were processed for histologic slide preparation in a Shandon Citadel 2000 (Thermo Scientific) tissue processor using standard techniques. After processing, samples were paraffin-embedded and cut to 5 µm sections. Sections were stained with haematoxylin-eosin (HE) in MICROM Auto stainer HMS 70 (Thermo Scientific), coverslip mounted by Neomount (Merck) and evaluated for patho histological changes using an Olympus CX40 microscope equipped with an Olympus Camedia C-4040 camera.
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