A New Challenge for Fluorescence Microscopy

Stem Cell Biology in Cancer Research

Contact and Information

Prof. Mario Faretta, European Institute of Oncology, Milan, Italy
Mario.Faretta@ifom-ieo-campus.it

Images and Videos: © M. Faretta

: 1: Two photon imaging of a MCF10A spheroid. MCF10A immortalized mammary epithelial cells were infected with a pBabe-GFP-H2B retroviral construct and grown in 3D culture conditions. Images were acquired using a Leica TCS SP5 confocal microscope coupled with a Chameleon Ultra II infrared pulsed laser tuned at 950 nm. A 63 glycerol immersion objective was employed thanks to its optimal numerical aperture (NA 1.3) and increased free working distance (0.28 mm).

: 2: Segregation of Numb protein during cell division in mammary stem cells. The spatial distribution of the polarity marker Numb (Green: Alexa 488; Cyan: DAPI) has been analyzed in normal (A) and cancer (B) stem cells. The 3D projection elaborated from a stack acquired with a Leica TCS SP2 confocal microscope clearly shows that the asymmetric partitioning typical of normal stem cells is lost upon transformation.

: 3: Two photon imaging of NAD(P)H in MCF10A mammary epithelial cells. Infrared pulsed excitation at 740 nm has been employed to excite fluorescence of endogenous molecules in in vitro grown mammospheres. The spectral analysis shown in the graph reveals the molecular sources contributing to the generation of the emitted light. Stacks were acquired using a Leica