Ultra-large field of view chip-based TIRF super-resolution microscopy for life sciences

Session

Session 1 - Multimodal Imaging including Correlative

Authors

Balpreet Singh Ahluwalia (1), Jean-Claude Tinguely (1), Luis E. Villegas-Hernández (2), Nikhil Jayakumar (1), Vishesh Kumar Dubey (1)

Affiliations

1. UiT, The Arctic University of Norway
2. UiT, The Arctic University of Tromsø

Keywords

Super-resolution microscopy

TIRF

Multi modal microscopy

High throughput 

Abstract text

Laser illumination engineering together with alteration of the photophysical properties of the fluorophores opened the opportunities of surpassing the diffraction limit of an optical microscope. Here, the optical microscopy body have gone thorough major improvement, while the sample holder made of glass slide or glass coverslips remained unaltered. In this talk, an overview of photonic chip-based multi-modal super-resolution TIRF optical microscopy platform is presented. Instead of a glass coverslips, the sample is seeded directly on top of an optical waveguide, (photonic-chip), that delivers the evanescent field illumination directly to the sample via total internal reflection (TIR). The core of chip is made of high-refractive index material ensuring excellent optical sectioning via ultra-thin (decay <50nm), ultra-large and clean TIR illumination over entire length of the chip (centimeter scale) and supports broad spectral range. A highly multi-moded optical waveguide is essential to generate uniform TIRF illumination over large areas and to avoid shadowing problem, an analogy to this can be drawn to ring-TIRF approach via objective lens. The photonic-chip is capable of simultaneously generating TIR illumination over all supported wavelengths, without the need of mechanical readjustments. 

The photonic-chip based microscopy not only reduces the footprint, and complexity but enables integration of different microscopy platforms such as on-chip single molecule localization optical microscopy (SMLM) [1], on-chip TIRF-structured illumination microscopy (TIRF-SIM) [2], light intensity fluctuation based optical super-resolution microscopy [3] and its compatibility with correlative light-electron microscopy [4]. The chip-based SMLM  enabled super-resolved images over millimetre field-of-view scale; a 100-fold increase in imaging area as compared to conventional SMLM platforms, thus opening the opportunities of high-throughput optical nanoscopy. The compatibility of photonic-chip for different biological applications have been demonstrated on living  (5) and delicate cells such as neurons (6). Similarly, the photonic-chip withstands standard preparation protocols of histopathology (7). This makes photonic-chip optical microscopy an attractive platform for application looking for scanning large areas with super-resolution and ultra-high contrast. A short discussion will be presented on recent development to harness the dark-field alike TIR-illumination from a photonic-chip for label-free superior contrast imaging (8) and label-free super-resolution imaging (9) of nanosized extra-cellular vesciles and tissue sections.

References

1. R. Diekmann, O. I. Helle, C. I. Oie, P. McCourt, T. R. Huser, M. Schuttpelz, and B. S. Ahluwalia, “Chip-based

wide field-of-view nanoscopy,” Nature Photonics 11, 322 (2017).

2.. Ø.I. Helle, F.T. Dullo, M. Lahrberg, J.C.Tinguley, O.G. Hellesø and B. S. Ahluwalia, “ Structured illumination microscopy using a photonic chip. Nature Photonics 14, 431–438 (2020).

3. N. Jayakumar, Ø.I. Helle, K. Agarwal and B. S. Ahluwalia, “On-chip TIRF nanoscopy by applying Haar wavelet kernel analysis on intensity fluctuations induced by chip illumination”, Opt. Express, 28, 35454, 2020.

4. J.C. Tinguely, A. M. Steyer, C. I. Øie, Ø.I. Helle, F.T. Dullo, R. Olsen, P. McCourt, Y. Schwab, B. S. Ahluwalia, “Photonic-chip assisted correlative light and electron microscopy”, Communication Biology 3, 739 (2020).

5. J.C. Tinguely, Ø. I. Helle, and B. S. Ahluwalia, "Silicon nitride waveguide platform for fluorescence microscopy of living cells," Opt. Express 25, 27678-27690 (2017).

6. I. S. Opstad, F. Ströhl, M. Fantham, C. Hockings, O. Vanderpoorten, F. W. van Tartwijk, J. Q. Lin, J.C. Tinguely, F. T. Dullo, G. S. Kaminski-Schierle, B S. Ahluwalia, C F. Kaminski, “A waveguide imaging platform for live-cell TIRF imaging of neurons over large fields of view”, Journal of Biophotonics, 13, 6, e201960222, 2020.

7. Villegas-Hernández, L.E., et al., Chip-based multimodal super-resolution microscopy for histological investigations of cryopreserved tissue sections. Light: Science & Applications, 2022. 11(1): p. 1-17.

8. N Jayakumar, L E. Villegas-Hernández, W. Zhao, H. Mao, F. T Dullo, J.C Tinguley, K. Sagini, A. Llorente, B. S. Ahluwalia, “Label-free incoherent super-resolution optical microscopy”, arXiv:2301.03451”, 2021.

9. N. Jayakumar, F.T. Dullo, V. Dubey, A. Ahmad, F. Ströhl, J. Cauzzo, E. M. Guerreiro, O. Snir, N. Skalko-Basnet, K. Agarwal, B. S . Ahluwalia, "Multi-moded high-index contrast optical waveguide for super-contrast high-resolution label-free microscopy" Nanophotonics, vol. 11, no. 15, 2022,.