Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience
Article Title: Identification of a Novel Oligodendrocyte Cell Adhesion Protein Using Gene Expression Profiling
Figure Lengend Snippet: Oligodendrocytes were purified directly from the whole brain of postnatal day 7 rats using flow cytometry. A , FACS density scatter plot in pseudocolor shows forward scatter (FSC) versus side scatter (SSC) properties used to identify viable cells (gate R1). B , A2B5 and O4 immunofluorescence signals were used in combination with gate R1 to sort-purify homogeneous populations of vital cells at early and later stages of oligodendrocyte differentiation. Gates were set to collect single A2B5 + progenitors expressing middle to high levels of A2B5 (top left quadrant, R2) and single O4 + oligodendrocytes (bottom right quadrant, R3). C , Agilent Bioanalyzer analysis of total RNA extracted from oligodendrocytes isolated by FACS demonstrates the intact 28S and 18S rRNA bands. D , Sorted A2B5 + cells cultured for 6 d differentiated into mature oligodendrocytes, as evidenced by strong immunoreactivity with the O1 antibody (red fluorescence). Scale bar, 10 μm.
Article Snippet: The quality of total RNA was assessed using a Bioanalyzer microchip (Agilent, Palo Alto, CA).
Techniques: Purification, Flow Cytometry, Cytometry, FACS, Immunofluorescence, Expressing, Isolation, Cell Culture, Fluorescence