Determination of sub-diffraction-limited organelles and biomolecules using microsphere lenses in combination with fluorescence microscopy

Hui Yang; Norman Moullan; Johan Auwerx; Martin A. M. Gijs

doi:10.1117/12.2078360

2 March 2015 Determination of sub-diffraction-limited organelles and biomolecules using microsphere lenses in combination with fluorescence microscopy

Hui Yang, Norman Moullan, Johan Auwerx, Martin A. M. Gijs

Author Affiliations +

Proceedings Volume 9328, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII; 932804 (2015) https://doi.org/10.1117/12.2078360
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

We propose the use of a high-refractive index glass microsphere in combination with a conventional fluorescence microscope for imaging sub-cellular organelles and biomolecules. The microsphere is placed on a sample that is immersed in water, collects the near-field nano-features of the sample and generates a magnified virtual image in the farfield, which is recorded through a conventional water immersion objective. We first investigate the imaging capability of the microspheres on size-calibrated fluorescent micro-/nano-particles. The experimental results obtained from a microsphere with 60 μm in diameter demonstrate imaging capability of features of ~λ/7-size (λ is the wavelength) with a magnification factor of 5.4. The position of the virtual image, the field-of-view (FOV) of the microsphere and the magnification factor are studied by using microspheres with different sizes. Finite Element Method (FEM) simulations are performed, providing key insight into the imaging effect of the microspheres with super-resolution capability. Moreover, the distribution and complex shape of different sub-cellular organelles, like centrioles, mitochondria and chromosomes in the AML12 cell line, are imaged with help of the glass microspheres. Thereafter, the subcellular location of mitochondrial encoded proteins could be studied.

Citation Download Citation

Hui Yang, Norman Moullan, Johan Auwerx, and Martin A. M. Gijs "Determination of sub-diffraction-limited organelles and biomolecules using microsphere lenses in combination with fluorescence microscopy", Proc. SPIE 9328, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII, 932804 (2 March 2015); https://doi.org/10.1117/12.2078360

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available