Crypt Plating
Crypt Plating - Related Videos
04:37
Intestinal Crypts Isolation: A Method to Isolate Whole Crypts from Small Intestine of Murine Model
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2023
In this video, we demonstrate a technique to isolate whole crypts from a mouse’s small intestine. The isolated whole crypts can be used to propagate long-lived and self-renewing 3D organoids in vitro.
13:29
Jet Impinging on an Inclined Plate
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2023
Source: Ricardo Mejia-Alvarez and Hussam Hikmat Jabbar, Department of Mechanical Engineering, Michigan State University, East Lansing, MI The goal of this experiment is to demonstrate how a fluid flow exerts forces on structures by conversion of dynamic pressure into static pressure. To this end, we will make a plane jet impinge on a flat plate and will measure the resulting pressure distribution along the plate. The resultant force will be estimated by integrating the product between the...
06:57
Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts
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Cited by 12 •
2021
In this protocol, we describe how to generate murine primary epithelial colon monolayers directly from intestinal crypts. We provide experimental approaches to generate confluent monolayers on permeable filters, confluent monolayers for scratch wound healing and biochemical studies, and sparse and confluent monolayers for immunofluorescence analysis.
10:53
Serial Dilutions and Plating: Microbial Enumeration
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2023
Source: Jonathan F. Blaize1, Elizabeth Suter1, and Christopher P. Corbo1 1 Department of Biological Sciences, Wagner College, 1 Campus Road, Staten Island NY, 10301 Quantitative assessment of prokaryotes can be onerous given their abundance, propensity for exponential proliferation, species diversity within a population, and specific physiological needs. Compounding this challenge, is the four-phase nature in which bacteria replicate (lag, log, stationary and death). The ability to accurately...
18:00
Aseptic Laboratory Techniques: Plating Methods
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Cited by 438 •
2012
When working with media and reagents used to culture microorganisms, aseptic technique must be practiced to ensure contamination is minimized. A variety of plating methods are routinely used to isolate, propagate, or enumerate bacteria and phage, all of which incorporate procedures that maintain the sterility of experimental materials.
FAQs
Crypt plating uses two complementary separation forces. Mechanical disruption breaks intestinal tissue into smaller fragments, while enzymatic digestion helps release crypt structures from surrounding tissue. This combination makes the crypts available for culture without relying on only one form of tissue processing. The isolated material can then be transferred into conditions designed to support subsequent organoid formation.
The extracellular matrix provides a supportive three-dimensional environment in which isolated crypts can organize, while defined growth factors supply signals that promote organoid formation. These components serve different functions: one provides structural support and the other regulates growth. Together, they create controlled culture conditions for studying intestinal epithelial behavior outside the original tissue.
Three-dimensional organoids preserve a tissue-like setting for investigating cells derived from intestinal crypts, including stem and progenitor cells. This format allows researchers to examine epithelial regeneration and tissue behavior under controlled laboratory conditions. Because the organoids originate from intestinal structures, they provide a model for studying biological responses that may be difficult to assess in isolated cells alone.
The workflow begins with intestinal tissue collection and preparation, followed by mechanical disruption and enzymatic digestion to release crypts. Researchers then place the isolated structures in an extracellular matrix and provide defined growth factors. Under these controlled culture conditions, the crypts develop into three-dimensional organoids that can be used for downstream medical and biological investigations.
Crypt plating supports studies of epithelial regeneration, intestinal disease, and host responses. Organoids generated from isolated crypts provide a controlled system for examining how intestinal tissue behaves under laboratory conditions. This makes the technique relevant to questions about tissue repair and disease-related changes, while keeping the experimental focus on intestinal epithelial structures and their responses.
The resulting organoids can be used in drug screening and toxicity studies, allowing researchers to evaluate responses in cultured intestinal tissue models. When the starting material reflects an individual patient, the system can also reveal patient-specific tissue behavior. These uses support research into personalized therapies and may reduce reliance on animal models during selected stages of investigation.