Some researchers prefer to include loops which are detected partially, others prefer to exclude loops which are only partially in the captured image. The ACAS default setting is to exclude partial loops. This can be overridden by adding this text in the experiment-set description:
For a three-dimensional collagen lattice with a final volume of 150 µl, the following protocol is recommended. The collagen lattice is prepared in two parts: 100 µl collagen preparation and 50 µl cell suspension
Part A. Collagen Preparation
5 µl bicarbonate (e.g. Sigma-Aldrich S8761-100ML)
For fluorescence, phase-contrast, bright field, and dark-field imaging, we recommend the following ibidi chambers. These chambers have high-resolution bottoms and chamber walls that will not interfere with the side-illumination used in dark-field.
About Plate Selection and Reports
Because the final report will compare results from all wells in the plate, the plate selection is important for creating the correct number of wells. For example, if you choose a 24 well plate, 24 wells will be created and the report will compare the results for 24 wells.
A 2D imaging experiment setup is preferred for un-labeled cell tracking of smaller cells such as immune cells. In MetaVi Labs' pathotaxis experiments when immune cells are co-cultured with pathogenic cells, 2D allows improved tracking of the immune cells (when the immune cells are un-labeled).
While tumor cells will adhere to glass or plastic, immune cells require a protein coating. Fortunately, this is easily accomplished with slides from ibidi.
Modifications to ibidi Protocols
We recommend ibidi slides and their associated protocols. However, please note: