eclipse ni e c2 confocal microscope  (Nikon)

Journal: Communications Biology

Article Title: Optogenetic control of integrin-matrix interaction

doi: 10.1038/s42003-018-0264-7

Figure Lengend Snippet: Validation of cell–matrix interaction with different cell lines stably expressing OptoIntegrin. a HEK-293T, HeLa and MCF7 cells stably expressing OptoIntegrin or moxGFP-PIF S were seeded on PhyB 1–651 -coated glass slides and incubated under 660 nm or 740 nm light (I = 20 µmol m −2 s −1 ) for 5 h and subsequently imaged using a transmission light microscope (scale bar = 200 µm). b Live cell imaging of cell–matrix interaction under 660 nm light and then switch to 740 nm light after 35 min with binary images of cell shapes to illustrate the cells spreading. For this, HeLa cells stably expressing OptoIntegrin were seeded on OptoMatrix and subsequently imaged. Micrographs were taken at indicated time points (scale bar = 10 µm). c Spatial control of cell attachment with OptoIntegrin-expressing HEK-293T cells. OptoIntegrin-expressing cells were cultivated on OptoMatrix, locally illuminated with 660 nm or 740 nm for 3 min and then left in darkness for 4 h. Afterwards, cells were fixed and imaged (scale bar = 200 µm)

Article Snippet: Finally, the stained coverslips were mounted on microscope slides with Mowiol 4-88 (Carl Roth, cat. no.: 0713), and confocal images were acquired on an upright confocal microscope (Nikon Instruments Eclipse Ni-E with a C2 confocal laser scanner, 100 × oil objective NA = 1.45 or 60× oil objective NA = 1.40).

Techniques: Biomarker Discovery, Stable Transfection, Expressing, Incubation, Transmission Assay, Light Microscopy, Live Cell Imaging, Control, Cell Attachment Assay