Differential Interference Contrast (DIC) microscopy is a label-free technique crucial for visualizing transparent biological specimens. In this study, we introduce a novel DIC interferometry imaging technique based on calcite beam displacer (CBD), significantly enhancing resolution and contrast. Our system, requires only two polarizers and a calcite beam displacer, offers a practical and straightforward implementation on standard microscopes. We demonstrate its efficacy by imaging human red blood cells (RBCs) and bacteria, retrieving differential phase information, and recording the dynamics of U2OS cells over an hour at 15 frames per second (fps). This advancement in DIC microscopy holds promise for broader applications in high-resolution imaging of transparent specimens in cell biology research.
Quantitative phase microscope (QPM) is used for the quantitative information and dynamic phase imaging of biological specimen, which provides wide application in biomedical sciences. High temporal phase stability of the QPM system is the primary requirement for accurate phase measurement. We have developed a common-path QPM geometry based on beam displacer and pinhole unit to achieve high temporal stability. The convenient adjustment of reference and object beams makes optical system compact and low-cost. The membrane fluctuations and qualitative phase are measured to demonstrate the capability and applicability of the system.
We report the development of field-portable multi-modal chip-based fluorescence, bright field and quantitative phase microscopy using smartphone detecting system. Fluorescence microscopy provide molecular information of the specimen with excellent specificity, while phase microscopy provides quantitative information of the specimen. Quantifying the optical phase shifts associated with biological structures gives access to information about morphology and dynamics at the nanometer scale. Here, we propose an integrated waveguide chip-based total internal reflection fluorescence (TIRF) microscopy and quantitative phase microscopy (QPM). We have developed microLED with cylindrical beam profile to couple excitation light into the edges of glass slide easily and efficiently. The evanescent field present on top of a waveguide surface is used to excite the fluorescence and a mobile phone microscope is used to collect the signal. Waveguide chip-based TIRF microscopy benefits from decoupling of illumination and collection light path, large field of view imaging and pre-aligned configuration for multi-color TIRF imaging. Light for bright field imaging and QPM integrated in the transmission mode. A microscope objective is used for collecting the fluorescence excited by evanescent field and transmitted light for bright field and quantitative phase microscopy (QPM). A compact and common path interferometer is used for QPM. The entire device is fabricated using three-D printer and integrated into one, which is compact and field portable. Images are recorded using a smart phone. Experimental results of onion epithelial cells, polystyrene microspheres and normal breast tissue are presented. The cost of entire system is very less.
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