| Female Investigators in Nonlinear Optical Nanoscopy

Fixed HeLa cells that have internalised 15nm AuNP via Tf-mediated endocytosis imaged by (a) Differential interference contrast (DIC) microscopy (b) co-circularly polarised reflection amplitude A⁺_2r (c) co-circularly polarised FWM amplitude A⁺_FWM (d) confocal fluorescence microscopy. FWM was acquired with pump-probe delay time of 0.5ps, pump (probe) power at the sample of 50μW (25μW). Confocal fluorescence and FWM are shown as maximum intensity projection of z stacks which were acquired over 6μm in 300nm z-steps, while the reflection is on a single xy plane.

Multiphoton imaging of human pancreatic islets-derived extracellular vesicles (antibody stain – yellow) embedded in COL1 hydrogels (SHG; purple). [less]

Multiphoton imaging of human pancreas tissue. Pancreas tissue was labeled with action (green) while the collagen network was imaged in a label-free way using SHG (purple). Detail imaging of the collagen fibrillar network at the right panel. [less]

Differentiated adipose-derived stem cell illuminated label-free with nonlinear microscopy - magenta: cellular lipid depots in adipocyt, yellow: intrinsic cellular fluorescence. (scale bar: 15 µm) [less]

Gonadal white adipose tissue illuminated label-free with nonlinear microscopy - cyan: collagen fibers, magenta: cellular lipid depots in adipocytes, red: red blood cells. (scale bar: 40 µm) [less]

Interscapular brown adipose tissue illuminated with nonlinear microscopy - magenta: cellular lipid depots in adipocytes, green: cellular mitochondria, extra-cellular elastin fibers. (scale bar: 10 µm) [less]

© Naya Giannakopoulou<br />Acknowledgements: Collaborators from the Cardiff School of Pharmacy and the Nottingham School of Pharmacy for the sample preparation and especially Dr. Paul Moody and Dr. J.Magnusson.

Example of background free heterodyne FWM imaging of 15nm polymer coated AuNPs uptaken in HeLa cells via Tf-mediated endocytosis.

30 x 30μm raster scan images of the reflection channel [a] and the corresponding FWM Amplitude channel [b] acquired with 140nm step size and 2ms dwell time.

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© Naya Giannakopoulou
Acknowledgements: Collaborators from the Cardiff School of Pharmacy and the Nottingham School of Pharmacy for the sample preparation and especially Dr. Paul Moody and Dr. J.Magnusson.

[a] Sketch of FWM epi-detected microscope with heterodyne interferometry implemented [b] Sketch of polymer coated 15nm AuNP. The polymer has attached fluorescently labelled ligand (Transferrin-AlexaFluor488) in order to follow the endocytic pathway inside HeLa cells. [c] DIC image of HeLa cells obtained at the microscope shown in (a) with a 100x, 1.45NA oil objective. FWM imaging was performed on the region drown with dashed rectangular [d] Set of 20x20μm raster scan FWM images show the distribution of the AuNPs in the cell region [e] Maximum Z intensity projection of the confocal fluorescent images indicating highly uptake efficiency of the fluorescently labelled AuNPs compounds.

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Darkfield image of 40 nm gold nanoparticles. Slightly larger particles look yellowish, while white spots are glass defects or other non-metallic scatterers. [less]