Journal: ACS nano

Article Title: Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy

doi: 10.1021/acsnano.5b03970

Figure Lengend Snippet: Live fluorescence microscopy identifies a region of interest for high-resolution SEM (a) Overlay of reflection and fluorescence images of CV1 cells on SiN membranes labeled with EGF-QDOTs (b) Time-lapse images of the white rectangular area in (a), which shows highly dynamic EGF-QDOT transport in the live-cell FM (see also Supporting Information Video S2). Some EGF-QDOT signals, like the one denoted with an asterisk, appear quasi-static while others are rather dynamic. The last panel shows the overlay of the higher magnification SEM images with the fluorescent image of the same regions. (c) SEM snapshot of the area depicted in (a) and (b). (d) Higher magnification image of the blue boxed area in (c). (e) Higher magnification image of the green boxed area in (c). A higher concentration of QDOT clusters is visible around the cellular cortex and at crossings of QDOT tracks. (f) Line profiles of the two EGF-QDOT complexes indicated with arrows in (d) and (e), with fwhm of 22 and 23 nm, respectively. (g) FM-SEM overlay of the red boxed area in (c) highlighting resolution difference between QDOT in FM (red) and EM contrast (black).

Article Snippet: Culturing CV1 Cells Monkey kidney fibroblast (CV1) cell line (Cell Lines Service, Germany), was maintained in DMEM supplemented with 10% fetal bovine serum, penicillin/streptomycin, and 2 mM glutamine at 37° under 5% CO 2 .

Techniques: Fluorescence, Microscopy, Labeling, Concentration Assay

Journal: ACS nano

Article Title: Electron Microscopy of Living Cells During in Situ Fluorescence Microscopy

doi: 10.1021/acsnano.5b03970

Figure Lengend Snippet: SEM snapshots of EGF-QDOT uptake. (a) Fluorescence image of CV1 cells on SiN membranes labeled with EGF-QDOTs. Live cell fluorescence microscopy observations highlight some regions as active sites for EGF(R) uptake. (b) Time-lapse series of the, white boxed area in (a); see also Supporting Information Video S3 imaged in the bionanoreactor in the integrated microscope with FM. Numbered regions appeared as highly dynamic uptake regions in FM observations, definig the region of interest for a SEM snapshot. The last panel of the time-lapse series shows the overlay of the higher magnification SEM images with the fluorescent image of the same regions. (c) Higher magnification overlay of fluorescent and SEM images of the area in the timelapse series depicted as 1. Similarly, panel (d) shows an overlay of fluorescent and SEM images of the labeled as 2. EGF-QDOT conjugates are visible as dark spots and localize especially around the plasma membrane and around filopodial structures.

Article Snippet: Culturing CV1 Cells Monkey kidney fibroblast (CV1) cell line (Cell Lines Service, Germany), was maintained in DMEM supplemented with 10% fetal bovine serum, penicillin/streptomycin, and 2 mM glutamine at 37° under 5% CO 2 .

Techniques: Fluorescence, Labeling, Microscopy