Optical microscopy is an invaluable characterization tool in biological, chemical, and materials sciences from both fundamental studies and applied viewpoints. There is a physical limitation imposed on the spatial resolution of an optical microscope (through Abbe's law) below which the optical features cannot be resolved. We demonstrate here the use of salt-based plano-convex microlenses (MLs) composed of high-refractive index (n, 1.47
Aim 1 : We plan to experimentally and theoretically investigate the physical parameters of MLs that affect the imaging quality including magnification, spatial resolution and contrast. In general, the size, shape, refractive index of MLs, and ML-specimen distance affect the physical parameters (focal length, focal spot size, spatial resolution, magnification and depth of focus) of the microlens. These studies are crucial for enhancing our understanding of interaction of light with MLs, and for optimizing the performance of MLs for imaging applications in life-, biomedical- and materials-sciences. Through this specific aim, we intend to experimentally and theoretically investigate the effect of ML dimension, ML curvature (size), refractive index, and ML-specimen distance on magnification and spatial resolution. The experimental results will be corroborated with calculations based on geometric optics and electromagnetic theory using ray tracing and Finite Difference Time Domain (FDTD) methods.
Aim 2 : Imaging using optimized microlens-based nanoscopy. We plan to utilize the optimized MLs to acquire super-high resolution using a conventional optical microscope. The knowledge gained in aim 1 on optimization of MLs for super- high resolution will be utilized for imaging nanoscale biological particles and nanolithographic fabricated nanopatterns. Our nanoscope will consist of three major components: a conventional optical microscope, a (or an array) microlens(es) and a piezoelectric scanning stage. For the proof-of-concept experiments, we will perform both fluorescence and bright- field imaging and scanning of protein and DNA nanoarrays, bacteriophage and viruses particles in the 70-150 nm dimension range. Ultimately, the proposed imaging tool would be useful for scientists, engineers, and clinicians for chemical and biochemical imaging, probing cellular dynamics and processes on or near cell surfaces, and biosensing applications.
The proposed work will develop an inexpensive nanoscope that can resolve biological features below 100 nm through the novel use of delinquent-salt microlenses. These microlenses would be potentially very useful for studying dynamic and static biological processes on or near the cell surface, probing biological particles and large biomolecules, and biosensing applications.
|Rajasekaran, Pradeep Ramiah; Zhou, Chuanhong; Dasari, Mallika et al. (2017) Polymeric lithography editor: Editing lithographic errors with nanoporous polymeric probes. Sci Adv 3:e1602071|
|Rajasekaran, Pradeep Ramiah; Sharifi, Payam; Wolff, Justin et al. (2015) Fabrication and characterization of non-linear parabolic microporous membranes. J Memb Sci 473:28-35|
|McNichols, Colton; Wilkins, Justin; Kubota, Atsutoshi et al. (2014) Investigating surface topology and cyclic-RGD peptide functionalization on vascular endothelialization. J Biomed Mater Res A 102:532-9|
|Dogra, Navneet; Li, Xuelian; Kohli, Punit (2012) Investigating ligand-receptor interactions at bilayer surface using electronic absorption spectroscopy and fluorescence resonance energy transfer. Langmuir 28:12989-98|