High throughput microplate assays to evaluate lipid inducing effects of biological agents on meibomian gland epithelial cells and possibly other lipid-producing cells.
Two spectrophotometric microplate assays with dual staining for either fluorescent Nile red (NR) plus 4,6-diamidino-2-phenylindole (DAPI) or non-fluorescent Oil red O (ORO) plus Crystal violet (CV) were applied and optimised to evaluate the lipid producing capacity of immortalised human meibomian gland epithelial cells (iHMGEC). The microplate assays offer a rapid, high throughput and objective measurement of the amount of lipids in iHMGEC (and potentially other lipid-producing cells) and can be used for screening the effects of biological agents or incubation changes on lipid production in cells in future studies.
Compared to the microscopic imaging method, all measurements of the microplate assays are recorded by the microplate reader, providing unbiased objective results. Microplate assays combined with microscopic observations allow both visualisation of lipid distribution within cells and an objective semi-quantitative estimation of lipid production.