It is well known for structured illumination microscopy (SIM) that the lateral resolution by a factor of two beyond the classical diffraction limit is achieved using spatially structured illumination in wide-field fluorescence microscope. In the state of art SIM systems, grating patterns are generally generated by physical gratings or by spatial light modulators such as digital micro mirrors (DMD), liquid crystal displays (LCD). In this study, using a combination of LCD and ground glasses, size controlled randomized speckle patterns are generated as an illumination source for the microscope. Proof of concept of using speckle illumination in SIM configuration is tested by imaging fixed BPAE cells.
Axicon lenses are conical prisms, which are known to focus a light source to a line comprising of multiple points along the optical axis. In this study, we analyze the potential of axicon lenses to view, image and record the object behind opaque obstacles in free space. The advantage of an axicon lens over a regular lens is demonstrated experimentally. Parameters such as obstacle size, object and the obstacle position in the context of imaging behind obstacles are tested using Zemax optical simulation. This proposed concept can be easily adapted to most of the optical imaging methods and microscopy modalities.
Spatially non-uniform illumination patterns have shown significant potential to improve the imaging. Recent developments in the patterned illumination microscopy have demonstrated that the use of an optical speckle as an illumination pattern significantly improves the imaging resolution at the same time reducing the computational overheads. We present a DMD based method for generation of digital speckle pattern. The generated digital speckle and uniform white light illumination are used as two illuminations to acquire images. The image reconstruction algorithm for blind structured illumination microscopy is used to get the high resolution image. Our approach does not require any calibration step or stringent control of the illumination, and dramatically simplifies the experimental set-up.
Time averaged imaging is one of the widely used methods to achieve improved image quality, used in different types of microscopic methods. Time averaged imaging refers to adjusting the exposure time of the imaging system to obtain optimal images. In state of the art microscopes, the region of interest (ROI) of illumination beam for time averaged imaging can be selected to be of regular shapes such as circle or rectangle. This forces smallest possible ROI to be larger than the actual sample’s ROI which can be of a specific shape with complex contours. In this context, we present a flexible fiber bundle based illumination probe capable of illuminating samples of irregular shapes for time averaged imaging. Further, this probe is capable of multi-wavelength illumination, hence can be used for multi-fluorescence imaging. The fiber probe with features such as region selective and multi- wavelength illumination allows it to be used for optimal imaging of multi-fluorescence sample.
Imaging of physically inaccessible parts of the body such as the colon at micron-level resolution is highly important in diagnostic medical imaging. Though flexible endoscopes based on the imaging fiber bundle are used for such diagnostic procedures, their inherent honeycomb-like structure creates fiber pixelation effects. This impedes the observer from perceiving the information from an image captured and hinders the direct use of image processing and machine intelligence techniques on the recorded signal. Significant efforts have been made by researchers in the recent past in the development and implementation of pixelation removal techniques. However, researchers have often used their own set of images without making source data available which subdued their usage and adaptability universally. A database of pixelated images is the current requirement to meet the growing diagnostic needs in the healthcare arena. An innovative fiber pixelated image database is presented, which consists of pixelated images that are synthetically generated and experimentally acquired. Sample space encompasses test patterns of different scales, sizes, and shapes. It is envisaged that this proposed database will alleviate the current limitations associated with relevant research and development and would be of great help for researchers working on comb structure removal algorithms.
There was a renewed interest, during the recent years, in the imaging and tracking of targeted cells or organelles for
a variety of biomedical and lab-on a chip applications that include particles movement. However, nonspecific
illumination during tracking can have adverse effects such as heating, reduced image contrast and photo bleaching.
In fact, current available tracking and imaging systems are unable to selectively illuminate the particle being
tracked. To fill this void, we have developed a fiber optics based probe system incorporating a spatial light
modulator (SLM) and an imaging fiber bundle for selective illumination on the targeted particle. A GRIN lens is
attached at the distal endface of the image fiber bundle for optimised illumination and collection. A tracking
algorithm is developed in order to enable controlled illumination through SLM to target the illumination point or
location in accordance with the particle movement and size variation. Further with this probe, particles can be
illuminated with light pulses of controllable duty cycle and frequency. The proposed methodology and developed
probe have good significance and expected to find potential applications areas such as optogenetics, cell signalling
studies, and lab-on a chip systems.
Flexible fiber optic imaging systems including fiber optic confocal probes have found tremendous significance in the recent past for its applications in high resolution imaging. However, motorized stage is required for scanning the sample or tip of the fiber in fiber based confocal probes. In this context, we propose a fiber probe confocal system using digital spatial light modulator devoid of using a mechanical scanning stage. Each fiberlet in the image fiber acts not only as a light conduit but also as a confocal pinhole. The paper also introduces the variation in the contrast by varying the number of illuminated fiberlets which effectively implies variation in the effective pinhole size. This approach has enabled the probe to act as an imaging unit with resolution that can be controlled and varied from a wide-field to a confocal.
Visual access to physically inaccessible parts has become the forefront of research and development in medical diagnostics tools and procedures. Flexible and thin endoscopes with fiber bundle as an image conduit serves this purpose. However, when the light passes through the core of the fiberlet, it is blocked by the inter fiberlet gap. This structural limitation creates special honeycomb like pattern overlaying the image captured with the image fiber assisted probes, known as the comb structure or fiber pixelation. It obstructs the perception of the original image sacrificing resolution and contrast and inhibits the use of object recognition and tracking algorithms. Generally, comb structure removal or depixelation methods are employed to remove honeycomb pattern from an image. In the recent past, several depixelation techniques have been proposed albeit using different set of pixilated images by different researchers. It is quite difficult to make a comparison of their performances based on such images, as they adopt different images for different particular framework of their study. In this context, a basic database of such images is the need of the hour to meet the growing diagnostic needs in the medical and industrial arena. This paper in this context proposes and details a Comb Structure Affected Image database (CSAI) to meet the objective. Images are generated considering the image fiber specifications and the characteristics at different targeted optical imaging modalities delineated by resolution scales. The proposed database is designed to have a set of synthetically generated pixelated images of test patterns of different scales, sizes and shapes.
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