For injection, EdU (Carbosynth Limited) was resuspended at 5 mg/mL in saline and passed through a 0.2 μm filter. One mg was given by intraperitoneal (IP) injection. For continuous administration, mice were given an IP injection at the start of the experiment and then provided with EdU in the water, resuspended at 0.5 mg/mL with 1% sucrose and passed through a 0.2 μm filter. Water bottles were shielded from light and the water supply changed every three days. Samples were dissected and processed as for IF. For EdU detection, the Click-iT EdU Alexa Fluor 488 Imaging Kit (ThermoFisher Scientific) was used. For combined EdU and protein detection, EdU detection was completed first and then the IF protocol was followed beginning at the blocking step. Whole-mounts in which EdU was detected were acquired in a single frame with z-stack on the Nikon AZ100 macro confocal microscope and processed, including with maximum projection, with Fiji (ImageJ). Sections were imaged as described above.
Az100 macro confocal microscope
Manufactured by Nikon
The AZ100 macro confocal microscope is a versatile imaging tool designed for high-resolution, large field-of-view imaging. It features a wide magnification range, enabling detailed observation of specimens from small to large. The AZ100 utilizes confocal technology to provide optical sectioning and improved image quality, making it suitable for a variety of applications in research and industrial settings.
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Lab products found in correlation
2 protocols using az100 macro confocal microscope
1
EdU Labeling and Imaging Protocol
2
Fluorescent Imaging of Tympanic Membranes
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Fluorescent imaging of whole-mount TMs was performed on a Nikon A1R HD confocal microscope with a DU4G filter-based detector, using a Plan Apo Lambda 10× 0.45NA or Super Plan Fluor LWD 20x 0.70NA air objective lens with digital zoom of either 1x or 2x. TMs from K5-CreERT2; R26R-Confetti mice were acquired using a Nikon AZ100 macro confocal microscope with a DUS spectral detector using a 4x objective and 2x optical zoom as well as a 1× or 2× digital zoom. Both microscopes used NIS-Elements software for acquisition. Whole-mount images are displayed as maximum intensity projections of z-stacks. TM sections were imaged on a Leica DM6 B microscope. FIJI (ImageJ) software68 was used to analyze images, place scale bars, export individual TIFFs, and adjust levels for each channel as needed to maximize image clarity. Imaging of gross anatomy of TMs was performed using a Leica M205 FA stereo microscope and a 2× air objective with the LAS X software.
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