Organ Model: Colon

Version 1.0 (Release Date: March 2022)

Introduction

These Compound Distribution Kit analysis tools simplify assay preparation and data analysis for the Emulate Compound Distribution Kit (CDK) protocol, enabling users to quantify the amount of test compound loss (e.g., absorption or adsorption) ahead of a Chip-S1^® Stretchable Chip experiment.

There are two Excel-based tools in the download: The Media Volume Preparation Calculator and the CDK Data Analysis Calculator. The Media Volume Preparation Calculator calculates the media volume required for the CDK assay, based on the intended flow rate, study duration, and expected re-dosing protocol. The CDK Data Analysis Calculator calculates the adjusted average cellular exposure and concentration range based on the dosing concentration.

Related Information:

• Protocol: Compound Distribution Kit Protocol
• Protocol: Compound Treatment Solution Preparation and Treatment

Version 1.0 (Release Date: November 2021)

Introduction

The Apparent Permeability (P_app) Calculator is an Excel-based tool that helps users calculate the permeability of the epithelial-endothelial barrier in Emulate Organ-Chips. The P_app of a molecule is determined by dosing the inlet of one channel, collecting the effluent of both channels, and calculating the amount of compound that crossed through the membrane over time. This tool can be used to help researchers streamline the process of assessing the barrier function of healthy or diseased tissues, as well as studying drug transport.

Related information:

• Product Pages: Colon Intestine-Chip, Duodenum Intestine-Chip, Proximal Tubule Kidney-Chip
• Protocols: Barrier Function Analysis
• Technical Note: Characterization of the Colon Intestine-Chip

Version 1.0 (Release Date: June 2022)

Introduction

The Immune Cell Recruitment Analysis Tool allows researchers to quantify the migration of immune cell across the channels of Emulate Organ-Chips, such as the Colon Intestine-Chip. This tool is comprised of two solutions, an ImageJ2/Fiji script and a Python script. Using inputted 3D image z-slices, this Emulate solution performs 3D segmentation and spatial analysis of migratory immune cells.

Related Information

• Product Page: Colon-Intestine Chip
• Related Protocol: Basolateral Recruitment of Peripheral Blood Mononuclear Cells (PBMCs) on the Colon Intestine-Chip
• Application Note: Modeling Inflammation-Specific Immune Cell Recruitment in the Colon Intestine-Chip
• Webinar: Modeling Inflammatory Immune Cell Recruitment and Response with the Colon Intestine-Chip

Introduction

This document provides guidance for using the Pod Imaging Adapter, which organizes Pod Portable Modules and aligns them on the same viewing axis for quick cell viability and health checks.

Introduction

This is a revised version of the previously released “Basic Organ-Chip Culture” protocol (updated as of 22-Dec-2022)

This protocol covers the general process for using the Basic Research Kit.

Introduction

This protocol describes how to induce the migration of peripheral blood mononuclear cells (PBMCs) from the vascular (bottom) channel to the epithelial (top) channel in the Emulate Colon Intestine-Chip and assess downstream effector function.

RELATED information:

• Related analysis tool: Immune Cell Recruitment Analysis Tool

Introduction

This protocol uses the Primary Colon Intestine-Chip as a reference point aiming to the semi-quantitative assessment of the phosphorylated Signal transducer and activator of transcription 3 (STAT3) levels under baseline conditions and upon basolateral stimulation with the cytokine interleukin 22 (IL-22). These methods and assay conditions could change with a different Organ-Chip or stimulant.

Introduction

This is the method we have developed for fixation and immunofluorescence staining for the Colon Intestine-Chip. We realize, however, that users may have their own fixation and staining processes that have been developed for specific cell types, antibodies, or antigens. If users would prefer to use other fixatives, permeabilizing solutions, or blocking buffers, they may do so while following the process outlined below.

Introduction

This is the method for isolation and purification of the total RNA for the epithelial and endothelial cells of the Colon Intestine-Chip. For isolation and purification of customer originated cell material, this protocol might need to be optimized. Please reach out to Emulate Field Science Support for additional guidance.

Introduction

This is the method to identify the total amount of protein in epithelial cells lysate and/or effluent samples of the Colon Intestine-Chip. For protein quantification of customer originated cell material, this protocol may need to be optimized. Please reach out to Emulate Field Science Support for additional guidance.