Organoid Imaging and Analysis Protocol

Images of organoid cultures were captured using a Keyence BZ-X800 microscope with BZ-X800 Viewer software, v01.02.03.02 or an EVOS FL Auto System (ThermoFisher). Size and morphology of organoids was analyzed using OrganoSeg software^{52 (link)} and phase contrast images. Measurements were performed on digital images by blinded investigators using ImageJ software. For histological analyses, organoids were recovered from the culture plates and treated with Histogel (ThermoFisher) prior to formalin fixation and paraffin embedding, following standard protocols. Slides were stained with hematoxylin/eosin and with Alcian Blue to visualize mucus production. TEM was performed as previously described for human organoids^{87 (link)}. Briefly, organoids were fixed in 3% glutaraldehyde, processed for ultrathin sectioning, and then were imaged on a Zeiss LEO 912AB TEM.

Free full text: Click here

Hashimi M., Sebrell T.A., Hedges J.F., Snyder D., Lyon K.N., Byrum S.D., Mackintosh S.G., Crowley D., Cherne M.D., Skwarchuk D., Robison A., Sidar B., Kunze A., Loveday E.K., Taylor M.P., Chang C.B., Wilking J.N., Walk S.T., Schountz T., Jutila M.A, & Bimczok D. (2023). Antiviral responses in a Jamaican fruit bat intestinal organoid model of SARS-CoV-2 infection. Nature Communications, 14, 6882.

Publication 2023

EVOS FL Auto System Histogel LEO 912AB TEM

Alcian blue Eosin Formalin Glutaraldehyde Human Microscope Mucus Organoids Phase contrast

Corresponding Organization :

Other organizations : Montana State University, University of Arkansas for Medical Sciences, Colorado State University

Top 5 similar protocols

Variable analysis

independent variables

None explicitly mentioned

dependent variables

Size and morphology of organoids
Mucus production

control variables

None explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!