Journal: ACS biomaterials science & engineering

Article Title: Spatially Defined Cell-Secreted Protein Detection Using Granular Hydrogels: μ GeLISA

doi: 10.1021/acsbiomaterials.2c01308

Figure Lengend Snippet: (a) Representative maximum intensity projection of z-stack images of microgels incubated in different concentrations of MMP2. (i) 0, (ii) 300, (iii) 1200 ng/mL of MMP2. Purple: anti-MMP2. Green: microgel. Light blue: phalloidin 555. Scale bar: 50 μm (b) standard curve formed by measuring the average fluorescence of the microwells after being incubated at 0, 75, 150, 300, 600, and 1200 ng/mL. The measurements were then fit into a 4-parameter log-logistic regression. The curve has an inflection point at 457.28 ng/mL. Shaded area indicates 95% confidence interval. (c) Center z-slice of HSC spheroids with MMP2 μGeLISA. Purple: anti-MMP2. Light blue: phalloidin 555. Scale bar: 50 μm (d) average radial profile of MMP2 concentration around HSC aggregates of 10 and 50 cells. Shaded area shows the 95% confidence interval. n = 3 experimental replicates. (e) Total amount of secreted MMP2 by the aggregates within a 150 μm radius. Student’s t-test. *p < 0.05. n = 24–30 biological replicates.

Article Snippet: Recombinant human IL-6 protein (cat. no. 206-IL), human/primate IL-6 monoclonal antibody (dAB, cat. no. MAB206), biotinylated human/primate IL-6 (cAB, cat. no. BAF206), human MMP2 monoclonal antibody (MMP2 dAB, cat. no. MAB902), and human MMP2 biotinylated antibody (MMP2 cAB, cat. no. BAF902) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Incubation, Fluorescence, Concentration Assay