The Applied Nano-Optics group @ INL is developing new optical technology contributing to answer outstanding questions in the life sciences. We aim to develop and apply a variety of microscopic imaging tools to study lipid bilayers, liposomes and nanoparticle-lipid complexes within a biological native environment in real-time. Also, we actively promote the introduction of our research ideas into the market place through the protection of our IP.
Developments introduced by our group include a new type of combined optical and scanning probe microscopy  and advanced ellipsometry application for investigations at the solid-liquid interface [2,3]. Furthermore, we are actively developing a comprehensive framework for characterizing white light scattering of cellular exo- and endocytosis during microscopic observation [4,5].
The widespread availability of a variety of medical imaging technologies has constituted one of the driving forces behind the revolution in medical diagnosis over the last century. As of today, disease diagnosis often results from a combination of medical examinations that present a multiplexed view of the patient’s medical condition to the medical doctor. Similarly, the development of new medical treatment procedures has benefitted and will profit greatly from the widespread availability of new advances in microscopy that provide crucial new insights into human cell biology. These advances are spread over the realms of optical, electron and Scanning Probe Microscopy (SPM).
The Applied Nano-Optics Laboratory team has developed a new variant of combined microscopy, based on a fluorescence optical sectioning microscopy module and atomic force microscopy (AFM), a well-known variant of SPM. By creating a time independent fluorescence excitation light illumination of the AFM cantilever, we enable for the first time the simultaneous recording of fluorescence optically sectioned images and various AFM recording modes. We envision that this development will have a significant impact in the study of live-cell signalling processing in cellular biology and are currently exploring this with the team of Prof. Sandra Paiva at the University of Minho.
- Miranda A, Martins M, De Beule PAA. Simultaneous differential spinning disk fluorescence optical sectioning microscopy and nanomechanical mapping atomic force microscopy. Rev Sci Instrum. 2015; 86(9):093705.
- Miranda A, Martins M, De Beule PAA. Simultaneous Advanced Microscopies for Live Cell Signaling Dynamics Investigations. Biophys J. 2016;110(3):517a.
- Rosana Alves, Daphné Dambournet, Alexander Sorkin, Adelaide Miranda, Pieter A. A. De Beule, David Drubin and Sandra Paiva. Characterization of intracellular trafficking of nutrient transporters using combined fluorescence optical sectioning and nanomechanical mapping atomic force microscopy in mammalian cells. XIX. Annual Linz Winter Workshop. 3-6th February 2017. Advances in Single-Molecule Research for Biology & Nanoscience, Linz, Austria.
People at the project:
INL: Pieter De Beule, Marco Martins Adelaide Miranda
Scientific Visitors: Sandra Paiva, Rosana Alves, Claudia Barata Antunes
Endocytosis, the taking in of matter by a living cell by invagination of its membrane to form a vacuole, and exocytosis, the process of vesicles budding off membranes for the transport of membrane bound secretory vesicles to the extracellular matrix, exemplify fundamental processes in biology. In-vivo detection of both exo- and endocytosis currently represents a formidable challenge to image capture technology due to the speed of the event, the size of the lipid vesicles and optical contrast available.
This project aims to introduce an exact electromagnetic theoretical model for the quantitative understanding of how exo- and endocytosis is observed with video Differential Interference Contrast (DIC) microscopy.  In particular, it focuses on the fundamental understanding of how lipid-induced optical anisotropy influences the microscopic observation of exo- or endocytosis. For this project we gratefully acknowledge aid received through the SCATMECH polarized light scattering software library developed by Dr. Thomas A. Germer at NIST.
- De Beule PAA. Surface scattering of core–shell particles with anisotropic shell. J Opt Soc Am A. 2014; 31(1):162.
- SCATMECH Project Home Page
- Dylan Marques, Adelaide Miranda, Ana G. Silva, Pieter A. A. De Beule. Optical scattering of cellular exo- and endocytosis. XIX. Annual Linz Winter Workshop. 3-6th February 2017, Advances in Single-Molecule Research for Biology & Nanoscience, Linz, Austria.
People at the project:
INL: Adelaide Miranda, Pieter De Beule
Scientific visitor: Dylan Marques, Ana Silva
Thin film fabrication and characterization forms one of the cornerstones of modern nanotechnology. Optical models of these thin films often require more complexity than an isotropic model in order to describe device application. At INL we have introduced a new variant of spectroscopic imaging ellipsometer that delivers improved thin film anisotropy measurements on a microscopic scale with strongly improved performance at the solid-liquid interface. We have applied this system for the spatially resolved determination of lipid bilayer optical anisotropy and are currently exploring applications for the characterization of exotic new 2D materials including di- and trichalcogenides
We have also introduced advanced ellipsometry for the detection of protein adhesion to nanoparticles deposited on a substrate. Thereby, we propose ellipsometry as an alternative to standard Surface Plasmon Resonance detection platforms for monitoring protein surface adhesion.
- Miranda A, De Beule PAA. Microscopic thin film optical anisotropy imaging at the solid-liquid interface. Rev Sci Instrum. 2016;87(4):043701
- Viegas D, Fernandes E, Queirós R, Petrovykh DY and De Beule PAA. Adapting Bobbert-Vlieger model to spectroscopic ellipsometry of gold nanoparticles with bio-organic shells. Biomed. Opt. Express 8, 3538-3550 (2017)
People at the project:
INL: Pieter De Beule, Adelaide Miranda