Quick Start Guide
Detailed Instructions
Sterilize
Soak biopsies in 75% ethanol for > 3 hours. It is normal for the biopsies to float until fully saturated with EtOH.
Rinse biopsies 3x in sterile water.
Once sterile, biopsies can be stored at 4°C in dH₂O or PBS until ready for use.
Biopsies should be incubated in growth media prior to cell seeding to limit osmotic shock.
Cell Seeding
We recommend the Excell™ Scaffold biopsies are initially seeded in a cell growth media suspension between 50-100 uL. Cells are generally seeded at densities between 1E4 and 10E6 cells/scaffold biopsy. Multiple seeding events can be performed to increase cell density over 72 hours. Allow the cells to adhere for 6 hours before adding additional growth media to submerge the biopsy.
Microscopy
Excell™ Scaffolds are autofluorescent under FITC-like channels. Scaffolds can also be visualized using Calcofluor White, or Congo Red stains. Bromophenol blue is a useful scaffold counterstain for bright field microscopy.
Fluorescent stains such as Hoechst 33342 and DAPI are used to identify the nuclei of adhered cells. Phalloidin stains can be used to detect actin filaments (F-actin) associated with adhered mammalian cells.
Survey
Your data helps our data. We thank you for testing our Excell Scaffold in your labs and your conclusions help feed into a better understand of our Beta 1.2 and how we can improve your experience. For filling out the Experience Survey we offer you your next Culture Kit for free.