Wound healing and migration assays are done by seeding cells into the Culture-Insert 2 Well. After cell attachment, a cell-free gap is created in which the cell migration can be visualized.
Co-Cultivation / Invasion
The Culture-Insert 2 Well can also be used for seeding two different cell types. After the removal of the insert, the cell fronts can be analyzed for their invasional behavior.
Visualization of Cell Migration
Live cell imaging of a wound healing and migration assay using the MCF7 cell line. Objective lens 10x, phase contrast microscopy.
Culture-Inserts vs. Scratch Assay
At first glance, the methods of scratching and placing a Culture-Insert seem to be two very similar approaches to create a cell-free gap. However, at a closer look, these two methods differ in important aspects that could influence the outcome of the assay:
Cell seeding into designated areas
Scratching the cell surface with a needle or pipet tip
Varying (i.e., not reproducible)
Gap surface residues
Extracellular matrix residues possible
Vessel surface damage
Yes, due to mechanical scratching on the surface
Yes, due to scratching the cells
Segregation & signaling of necrotic/apoptotic cells
Internal reference *
* The internal reference addresses the question of whether or not two opposite cell fronts influence each other. By using the Culture-Inserts, it is possible to measure the speed of:
A: a cell front that is opposite another cell front; and B: a single cell front that does not have an opposite cell front.
Time-dependent changes of the open area due to cell migration. A comparison between gap creation by using the ibidi Culture-Insert 2 Well (left) and by scratching with a pipet tip. Data provided by M. Börries, University Freiburg, Germany.
Ali Alhoshani, King Saud University, Riyadh, Saudi Arabia
“The ibidi Culture-Insert 2 Well 24plate was unbelievably helpful when I wanted precise and consistent results for wound healing assay. It saved my lab so much time and effort. The web-based quantitative image analysis feature is a huge plus. Thank you for such useful products. "
Ali Alhoshani King Saud University Department of Pharmacology and Toxicology Riyadh Saudi Arabia
Henrike Rebl, University Medicine, Rostock, Germany
"I have tested the Culture-Insert 2 Well and 3 Well and they worked quite nicely. I will definitely use the ibidi Culture-Inserts and µ-Dishes again because they are so easy to use and they work great for fluorescence microscopy."
Gisa Gerold, TWINCORE – Center for Experimental and Clinical Infection Research, Hannover, Germany
“We tested the Culture-Inserts 2 Well and now use them successfully on human hepatoma cells (Lunet N3 hCD81 cells, which are derived from Huh-7.5 cells). We are very satisfied with the ibidi products and with the fast delivery of the test chambers.”
Gisa Gerold TWINCORE – Center for Experimental and Clinical Infection Research Hannover, Germany
Renate Gehwolf, Paracelsus Medical University, Salzburg, Austria
"We are currently using the Culture-Insert 2 Well to perform wounding experiments with somatic stem cells, and also the 2D chemotaxis slides to study the migratory behavior of these cells. We are very happy with the products, because they are user-friendly, especially for wounding assays, as we do not want to do scratching because of the inevitable high variability. However, with the ibidi Culture-Inserts, we can perform highly reproducible experiments. We will definitely use them again.”
Renate Gehwolf Institute of Tendon & Bone Regeneration Paracelsus Medical University Salzburg, Austria
Judith Cathcart, Advanced Optical Microscopy Facility, University Health Network, Toronto, Canada
"The ibidi chambers (Culture-Inserts 2 Well) we used in our wound healing assays helped to keep the cell free area clean and consistent in size without damaging the ECM or leading edge of the cell fronts, making analysis of results more manageable and comparable. They're convenient and easy to use and their small size helps to conserve precious cells and culture materials. We'd certainly use them again.“
Judith Cathcart Advanced Optical Microscopy Facility University Health Network Toronto Canada
Jason W. Bjork, PhD, 3M Corporate Research Materials Laboratory, St. Paul, USA
“The ibidi Culture-Inserts 2 Well for wound healing assays provide a new line of testing for us. We wouldn't do traditional scratch assays at all because of the inherent variation, but with the new inserts we're able to provide many images to support our research. Our results are repeatable and reliable regardless of which researcher conducts the assay.”
Jason W. Bjork 3M Research Materials Laboratory St. Paul USA
Ross Flockhart, PhD, Stanford School of Medicine, Stanford, USA
"The ibidi cell migration Culture-Inserts 2 Well enabled me to perform highly reproducible and quantitative migration experiments. Since physical scratching isn't required, uniformity is maintained and adverse wounding responses are not an issue.
The online quantification tool WimScratch for the assay is also fantastic and it yields quantitative data from the images extremely quickly with little effort required. I'd definitely recommend it for migration studies.“
Ross Flockhart, PhD Stanford School of Medicine Department of Dermatology Stanford USA