Stem Cell Research Source

Fluorochrome-conjugated antibodies to pluripotent stem cell markers for bioimaging and flow cytometric analysis.

A Human embryonic stem cells (H9 ) were cultured in mTeSR™1 maintenance media (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, coming soon) pseudo-colored yellow, Oct3/4 Alexa Fluor® 555 (Cat. No. 560306, coming soon) pseudo-colored red, and SSEA-4 Alexa Fluor® 488 (Cat. No. 560308, coming soon) pseudo-colored green. Cell nuclei were counterstained using Hoechst 33342 pseudo-colored 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.

B Human embryonic stem cells (H9) were grown in 6 well culture plates coated with BD Matrigel hESC-qualified matrix and mTeSR1 media. Cells were dissociated into a single cell suspension and then fixed with 4% paraformaldehyde, followed by BD Perm/Wash buffer. Cells were incubated with the same cocktail of fluorchrome-conjugated antibodies used in the imaging experiments described above except that Oct3/4 PE (560186) was used in place of Oct3/4 Alexa Fluor® 555. Cells were run on a BD™ LSR II and data was analyzed using BD FACSDiva™ software.