BD Pathway™ Software

Embryonic stem cells cultured under feeder-free conditions remain pluripotent

Human embryonic stem cells (H9) were cultured in mTeSR1 maintenance medium (StemCell Technologies) on BD Falcon™ 96-well imaging plates (Cat. No. 353219) that were coated with BD Matrigel™ hESC-qualified matrix (Cat. No. 354277). Cells were fixed with 4% paraformaldehyde, followed by BD Perm/Wash™ buffer (Cat. No. 554723). Multicolor cell staining was performed with the following antibodies: Sox2 Alexa Fluor® 647 (Cat. No. 560302) pseudocolored yellow, Oct3/4 Alexa Fluor® 555 (Cat. No. 560306) pseudocolored red, and SSEA-4 Alexa Fluor® 488 (Cat. No. 560308) pseudocolored green. Cell nuclei were counterstained using Hoechst 33342 pseudocolored blue. Small images on the left show individual antibody staining. The larger panel on the right represents the merged image. The cells were imaged on a BD Pathway 435 bioimager using a 10x objective.

Phagosomal escape assay to quantify bacterial count

These confocal collapsed stack images (40X, 0.9 NA) were taken of macrophages infected with wild-type (top) and LLO-minus mutant L. monocytogenes (bottom). The images were pseudocolored, merged, and cropped. Macrophage actin is green, bacteria are red, and colocalized signals appear as merges of these color channels. Images on right show segmentation masks generated after Sub Object counting from images on the left. The upper right plot shows time-dependent colocalization coefficients of wild-type and LLO-minus mutant bacteria with actin and vice versa, respectively (n = 4 wells). The lower right plot shows time-dependent quantification of bacterial count per macrophage (n = 4 wells).

Representative Images Using BD Bioimaging Certified Primary Conjugated Antibodies

Pseudocolored merged images of nerve growth factor treated PC12 cells. Nuclei are pseudocolored blue in all images. Neurites were stained with: (A) Alexa Fluor® 488 conjugated anti β-tubulin antibody (green), (B) Alexa Fluor® 555 conjugated anti β-tubulin antibody (red), and (C) Alexa Fluor® 647 conjugated anti β-tubulin antibody (magenta). Images were acquired using a 20X objective (0.75 NA).