Optical Nanoscopy

ERC Starting Grant

European Union’s ERC Starting Grant (Nanoscopy, 2013)

European Union’s ERC Starting Grant (Nanoscopy, 2013)

Balpreet Ahluwalia’s project on “High-speed chip-based nanoscopy to discover real-time sub-cellular dynamics” was awarded with European Research Council prestigious ERC Starting Grant in 2013. This will support Ahluwalia and his new team (2 PhD and 2 Post-Doc) for the next 5 years. The sole criteria of selection for this esteemed funding are scientific excellence of the project and a proven track-record of the candidate. ERC Starting Grant is an attractive long term funding scheme, the competition is extremely tough with success rate of only around 9-10%. The funding from EU will enable the PI (Ahluwalia) to establish his own research group and providing adequate mechanism to become research leader. The project aims to develop a new field of research activities in the cross-roads of integrated optics and bio-medical imaging. The project aims to develop a novel optical nanoscopy with high resolution (50-100 nm) and fast imaging speed (25 Hz).

Optical nanoscopy has given a glimpse of the impact it may have on cell-biology and medical care in the future. Slow imaging speed and the complexity of the current nanoscopy techniques limits its use for living cells. The imaging speed is limited by the complicated bulk optics that is used in present nanoscopy. In this project, I propose a paradigm-shift in the field of advanced microscopy by developing optical nanoscopy based on a photonic integrated circuit. The project will take advantage of nanotechnology to fabricate waveguide-chip for illuminating the sample, while fast telecom optical devices will provide light switching. This will enhance the imaging speed of chip-based nanoscopy. This unconventional route will change the field of optical microscopy, as a simple chip-based system can be added to a normal microscope, rather than relying on complicated systems for super-resolution microscopy.

The proposed optical nanoscopy will be employed to discover the dynamics (opening and closing) of fenestrations (100 nm pores) present in the membrane of a living liver sinusoidal scavenger endothelial cell (LS-SEC). It is believed among the hepatology community that these fenestrations open and close dynamically, however there is no scientific evidence to support this hypothesis because of the lack of suitable tools. The successful imaging of fenestration kinetics in a live cell during this project will provide new fundamental knowledge about the fenestration dynamics, which will help in improved diagnosis and drug discover of liver related diseases.


Research Methodology and planned task

It is necessary to develop suitable waveguides as described in Task 1, which is divided into two sub-tasks 1.1 (chip-design) and 1.2 (fabrication). The implementation of optical nanoscopy is covered in Task 2, which is again divided into 2 parts (experimental and algorithm development). Task 3 describe the application of the proposed integrated nanoscope for live cell imaging, while Task 4 covers the extension of the proposed nanoscopy for 3-D and nonlinear studied. The project will start from 1st Feb 2014 and it will have 2 Post docs and 2 PhD working in close collaboration to accomplish this challenging task. The group will be cross-disciplinary, with competence on waveguides, cell-handling and super-resolution microscopy. The planned tasks and the responsible team member are also highlighted with each task. Ahluwalia will be involved in all aspects of the project, while the team members will also be benefited with existing local, national and international collaborators.

Task 1.1: Chip design (Post-Doc 1):

Candidate need to simulate and design single mode waveguide based on high refractive index waveguide material. The candidate needs to investigate the bending losses, taper design and interference fringes. Several simulation packages can be used like Clewin, Fimmwave, Comsol).

Timeframe: 10-12 months, 2014

Internal collaborator: Prof Olav Gaute Hellesø (Uni. of Tromsø)

Milestone: Simulation and optimization of the waveguides and the chip design, and one or more masks for photolithography.

Task 1.2: Fabrication and characterization of waveguides, (Post-doc 1)

The fabrication will be done at Optoelectronics Research Centre, University of Southampton, UK in collaboration with Prof James Wilkinson. A post-doc will spend 6 months (several visits) at the ORC, UK to fabricate waveguides and perform initial characterization, hosted as full group members. The waveguide material is tantalum pentoxide (Ta2O5). Post-doc 1 is responsible for waveguide fabrication, while PhD 2 might be trained in entire fabrication process.

Timeframe: 18-24 months, 2014-2015.

External collaborators: Prof James Wilkinson (Uni. of Southampton)

Milestone: Fabricated low-loss waveguide-chip


Task 2: Implementation of Optical Nanoscopy  (Post-doc 2):

Task 2.1: Experimental implementation (Post-Doc, PhD 1):

For this job, required optics, lasers, mechanics, camera, etc, will be be acquired and the set-up will be built. The control system (eg. Using microprocessor or with LabView to begin with) for the experiment will be made to synchronize the timing of laser excitation, optical switch, camera and objective lens holder for z-steps. The set-up will be tested with known samples such as polystyrene nanosphere and eventually for live cell imaging applications. Post doc 2 is responsible for this job, while PhD 2 and PhD 1 will also provide inputs to this task, as this the main topic of the project.

Timeframe: 18-24 months, 2016-2017

External collaborators: Prof Thomas Huser (Uni. of Bielefeld)

Internal collaborator: Prof Olav Gaute Hellesø (Uni. of Tromsø)


Task 2.2: Reconstruction Algorithm (PhD 2):

Structured illumination microscopy is a computation trick, where unresolvable information (high spatial frequency) is encoded to generate observable images (Moiré fringes). From these images, the unresolvable information is computationally obtained by separating, shifting and re-assembling the images and the information. The candidate needs to modify existing SIM reconstruction algorithm for the proposed chip-based optical nanoscopy. Continued improvement of SIM algorithm will be required and will be carried thorough-out the project.

Timeframe: 18-24 months, 2015-2017

External collaborators: Prof Thomas Huser (Uni. of Bielefeld)

Internal collaborator: Prof Robert Jessen (Uni. of Tromsø)

 Milestone: Fully operational optical nanoscopy suitable for live cell imaging


Task 3: Cell biology and imaging liver cells fenestration using optical nanoscopy (PhD 1)

The chip-based optical nanoscopy will be employed for a dedicated bio-imaging application for liver cells. It is necessary for a PhD candidate (in bio-physics) to be trained in cell-biology and isolation of liver cells, which is a highly specialized field. The candidate will be recruited rather early in the project (before optical nanoscopy is ready) to get essential training in cell biology The candidate will also be involved in building optical nanoscopy, adapting nanoscopy for live cell experiments by adding stage-top incubator to control temperature, CO2, etc. The set-up will be first tested and optimized by acquiring fluorescence images of simpler cell-lines such as Jurkat or HeLa before working on primary cells (LS-SEC). The candidate will also investigate cells health on waveguide-chip and optimize fluorescence staining.

Timeframe: 12-18 months, 2017-2018

Internal collaborator: Prof Peter McCourt and Prof Bård Smedsrød’s (Department of Medical Biology, Uni. of Tromsø)

Milestone: Imaging of liver fenestration in live cells and imaging of fenestration dynamics


Task 4: Extension of optical nanoscopy for 3-D and non-linear optical nanoscopy (PhD 2)

The candidate will investigate the feasibility of 3-D and nonlinear chip-based optical nanoscopy based on numerical simulation (Comsol, Fimmwave, Matlab). Depending on progress, a separate mask will be designed and waveguides will be fabricated. The experimental set-up, image acquistion, and the control system for this job will be similar to Task 2.1. However, the reconstruction algorithm (task 2.2) must be modified.

Timeframe: 18-24 months, 2014-2016

Internal collaborator: Prof  Olav Gaute Hellesø (Uni. of Tromsø)

External collaborator: Prof James Wilkinson (Uni. of Southampton) and Prof Thomas Huser (Uni. of Bielefeld)  

Milestone: Implementation of non-linear and 3-D chip based optical nanoscopy.



If you are interested in more information, collaboration, PhD- or MSc-projects, please contact Balpreet Singh Ahluwalia

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Last updated: 05.10.2022 08:55