SmartFlareTM is a reliable method for assessing mRNA expression in single neural stem cells

Figure 1 SmartFlare^TM detection in 3 d in vitro neural stem cells. A and B: The expression pattern for both SmartFlare^TM CD133 and Oct4 probes showed a diffuse and spotted-like fluorescence from the perinuclear area to the peripheral cytological processes; C: SmartFlare^TM probe for Actin showed a robust and overall localization of red fluorescence as indicative of positive mRNA expression; D: Scramble probe-treated cells were almost completely lacking in unspecific red background. Nuclei were stained by Hoechst dye. Scale bar: 50 μm.

Figure 2 SmartFlare^TM detection in 7 d in vitro neural stem cells. A and B: SmartFlare^TM CD133 and Oct4 probes fluorescence was less intense than the signal detected in 3 d in vitro neural stem cells, but still present in almost all cells; C: SmartFlare^TM probe for Actin showed a robust and overall localization of red fluorescence as indicative of positive mRNA expression; D: Scramble probe-treated cells showed a very faint unspecific red background. Nuclei were stained by Hoechst dye. Scale bar: 50 μm.

Figure 3 SmartFlare^TM detection in 15 d in vitro neural stem cells. A and B: SmartFlare^TM CD133 and Oct4 probes showed a dramatic fluorescence downregulation that was limited to small cytoplasmic granules in less than half of the observed cells; C: SmartFlare^TM probe for Actin showed a robust and overall localization of red fluorescence as indicative of positive mRNA expression; D: Scramble probe-treated cells were almost completely lacking in unspecific red background. Nuclei were stained by Hoechst dye. Scale bar: 50 μm.

Figure 4 SmartFlare^TM detection in 30 d in vitro neural stem cells. A and B: SmartFlare^TM CD133 and Oct4 probes showed few cells expressing a tiny granular pattern in the cytoplasmic domain; C: SmartFlare^TM probe for Actin showed an abundant red fluorescence in all observed cells; D: Scramble probe-treated cells were almost completely lacking in unspecific red background. Nuclei were stained by Hoechst dye. Scale bar: 50 μm.