High-resolution multi-modal imaging of sub-cellular structures with low numerical aperture objective
Somaiyeh Khoubafarin, Peuli Nath, Saloni Malla, Durgesh Desai, William D Gorgas, Amit K Tiwari, Aniruddha Ray
TL;DR
This paper introduces a cost-effective method to achieve high-resolution imaging of subcellular structures using low numerical aperture objectives, enabling affordable and portable multi-modal microscopy.
Contribution
The novel use of a 2D microlens substrate to enhance resolution and capture evanescent waves with low N.A. objectives.
Findings
Sub-diffraction-limited resolution (<400 nm) was achieved using a 0.25 N.A. objective with a microlens substrate.
The method enables simultaneous scattering, phase, and fluorescence imaging of breast cancer cells and nanoparticle uptake.
The approach improves light capture efficiency and resolution by collecting evanescent waves.
Abstract
Imaging of subcellular structures, which underpins many of the advances in biological and medical sciences, requires microscopes with high numerical aperture (N.A.) objectives which are costly, complex, requires oil immersion and have very limited field-of-view, typically covering a handful of cells. Here, we leverage a low N.A. objective to simultaneously capture scattering, phase, and fluorescence images of subcellular structures in breast cancer cells (BT-20) and observe nanoparticle uptake, with sub-diffraction-limited resolution (<400 nm with a 0.25 N.A. objective) utilizing a 2-dimensional (2-D) microlens substrate. High resolution labeled and label-free images of subcellular components is made possible by implementing a specific configuration, wherein the sample is placed in close proximity to the microlens substrate, which results in efficient collection of the rapidly decaying…
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Taxonomy
TopicsAdvanced Fluorescence Microscopy Techniques · Photoacoustic and Ultrasonic Imaging · Optical Coherence Tomography Applications