Apparatus and Method of Fluorescent Bench Top Optical Imaging System for Bio-Imaging Application

Abstract

Fluorescence imaging for preclinical studies and understanding pathological conditions has gained significant attention due to its essential role in biological detection and targeted drug delivery and drug discovery. However, the widespread use of commercial optical imaging systems is hindered by their high cost and limited accessibility. This research presents the development of a cost-effective tabletop optical imaging system. The system's efficacy was evaluated on various in-vitro samples, including carbon quantum dots (CQDs), serially diluted CQD samples, CQD gels, and fluorescent powder accumulations in phantoms. The ex-ovo technique experiment with chicken embryos highlighted challenges in injection techniques, while subsequent tests with microfluidic chips validated the system's accuracy and performance. The system was integrated with a low-light-detecting camera and optical filters ranging from 400nm to 650nm, demonstrated excellent fluorescence detection capabilities. Notably, the imaging system successfully differentiated between emission of fluorescent and non-fluorescent samples, underscoring its potential for preclinical applications. Key advancements include the elimination of autofluorescence, enhanced low-light detection, and precise fluorescence detection in microfluidic channels of 250 microns. The developed prototype model offers significant advantages in accessibility and cost-effectiveness, making it a valuable tool for drug delivery, medication research, biomedical and biotechnology studies. This research establishes an affordable optical imaging system that can be widely adopted in various laboratory settings, enhancing the scope and efficiency of preclinical research.

Specification

Description:The following specification particularly describes the nature of the invention and the manner in which it is performed:
[01] FIELD OF THE INVENTION
The present invention relates to a method and apparatus which has fluorescence imaging process along with preclinical studies and pathological analysis in biological detection used in targeted drug delivery and drug discovery.
[02] BACKGROUND OF INVENTION
The scientific community has shown significant interest in the development of optical imaging systems, leading to numerous studies focused on designing systems customized for specific applications. These systems are typically engineered to function for a singular purpose using optical components that are readily available without the need for specialized procurement. However, the high cost of commercially accessible optical imaging systems, which often incorporate high-end cameras, detectors, and intricate software controls, is a significant barrier for many laboratories. Recognizing the constraints imposed by these costly systems, researchers have sought to develop more economically viable approaches. Previous studies have emphasized the need for optical imaging systems that are more accessible and adaptable across diverse laboratory environments. By leveraging existing resources and refining the design and functionality, the invention is to democratize optical imaging technology and broaden its accessibility for scientific endeavors. Instead of relying on specialized and expensive components, alternative solutions are being explored to achieve comparable performance at a reduced cost. This shift in focus toward cost-effective solutions democratizes access to optical imaging technology, facilitating its widespread adoption and fostering innovation across various scientific disciplines.
The invention is characterized by a comprehensive development process, precise alignment using optical principles, versatility for various applications, and affordability and accessibility. By offering a unique and accessible solution, our in vivo optical imaging system opens new avenues for research and innovation, empowering scientists to make significant contributions to the field of biomedical imaging.
Design and develop a novel optical imaging system capable of capturing and analyzing injected fluorescence in biological samples. This involves integrating various components and visualizing software to achieve high-quality imaging for reliable data acquisition.
The optic system detects targeted fluorescence accumulation in specific regions of interest, providing precise identification of affected areas. This instrument is constructed using appropriate components such as concave lenses, long-pass filters, low-light detectors, and 20 integrated LED light sources of 4 different colours. Optical alignment is a crucial step in ensuring optimal performance and accurate imaging. The objective was to precisely align the components to achieve proper focus, illumination, and fluorescence detection.
To minimize external light interference, the system was enclosed in an acrylic box and coated with black-matte paint. This creates a controlled environment in which stray light is minimized, ensuring accurate measurements and reducing background noise during imaging.
To validate the performance and functionality of the optical imaging system. This involves conducting experiments using various fluorescent dyes, fluorescent-dye-induced phantom gels, dyes injected into imaging egg embryos, and microfluidic chip channels. All these sample spectra were collected using an Edinburgh spectrofluorometer to confirm the accuracy of the instrument's emission wavelength as proof of concept.
To assess the system's capability to accurately detect and capture fluorescence in different sample media and small-diameter fabricated channels with the lowest dye concentration, thereby ensuring its reliability and effectiveness.
[03] SUMMARY OF THE INVENTION
The invention offers significant advantages over existing technologies, primarily in terms of cost-effectiveness, high-quality imaging capabilities, and enhanced flexibility and control.
- Providing a cost-effective solution allows researchers with limited resources to access high-quality optical imaging technology without incurring exorbitant costs.
- The versatility of the developed system allows for a wide range of applications, including drug delivery analysis, tumor detection, and angiogenesis growth factor analysis, thus enhancing its utility in biomedical research and diagnostics.
The system's built-in optical components streamlined the imaging process, making it easier for researchers to achieve accurate results without the need for additional specialized equipment.
A key advantage of the invention is its ability to mitigate issues such as autofluorescence, which can distort imaging results using appropriate filters and imaging techniques.
Furthermore, the system overcomes challenges commonly associated with more advanced and expensive devices, such as small form factor limitations and the lack of integrated detectors, ensuring reliable and effective performance across various sample types and sizes.
Overall, the present invention represents a significant advancement in optical imaging technology, offering improved efficiency, cost benefits, and accessibility for researchers in biomedical research and diagnostic applications.
[04] OBJECT OF THE INVENTION
The proposed in vivo imaging system represents an advancement in optical imaging technology by providing a cost-effective alternative to current commercial products. The primary objective was to achieve a performance comparable to that of expensive imaging systems using accessible components and open-source software.
A key feature of the developed system is its ability to capture high-quality images of fluorescent samples at lower concentrations. This study addresses challenges such as autofluorescence, small form factor limitations, and the lack of integrated detectors commonly encountered in existing imaging systems.
Through extensive testing with various concentrations of samples, particularly focusing on nanosized contrast agents such as carbon quantum dots, the system demonstrated excellent imaging capabilities. This is evident in its ability to visualize injected fluorescent materials within phantom gels and microfluidic chips, indicating its potential for detecting and capturing microscale images of samples.
Overall, the invention presents a new use for existing technology by offering enhanced performance and accessibility in optical imaging for research and diagnostics.

[05] BRIEF DESCRIPTION OF THE DRAWINGS
The invention will be better understood and objects other than those set forth above will become apparent when consideration is given to the following detailed description thereof. Such description makes reference to the annexed drawings wherein:
Fig. 1. Block diagram of the proposed method
Fig. 2. CAD designs of developed system
Fig. 3. Developed bench top in -vivo optical imaging system
Fig. 4. Photoluminescence data with different concentrations (left) with images using filter ranges from 400nm-650nm(right)
Fig. 5. CQD gel strands imaged under 400nm-650nm filter
Fig.6. Different fluorescent color powders under UV excitation source-395nm(left), with different range of filters. A-Neon orange, B-Neon pink, C-Neon green, D-Violet,
E-Neon golden yellow, F-Neon yellow
Fig.7. 6 color fluorescent powders imaged under optic system with different range of filters under Blue excitation source of (400-450nm) A-Neon orange, B-Neon pink, C- Neon green, D-Violet, E-Neon golden yellow, F-Neon yellow.
Fig.8. Fluorescent powders imaged with different optic filters under green excitation source (500nm), A-Neon orange, B-Neon pink, C-Neon green, D-Violet, E-Neon golden yellow, F-Neon yellow.
Fig.9. Microfluidic chip for analyzing output of imaging system

[06] DETAILED DESCRIPTION OF THE INVENTION
The developed optical imaging system provides a cost-effective alternative to expensive commercial imaging systems. It uses easily accessible components that can be procured in laboratories, significantly reducing the overall cost of the system. This makes it more accessible to researchers and institutions with limited budgets.
Flexibility and Control: Unlike existing imaging systems that rely on automated functions, the present invention allows for the manual control of camera settings. This provides researchers with greater flexibility and control during image acquisition, enabling them to optimize the imaging parameters for specific experimental requirements.
Miniaturization: The present invention overcomes the limitations of bulky imaging systems by utilizing a compact and portable design. The system makes it easier to handle and maneuver, facilitating imaging in confined spaces or during in-vivo experiments.
Reduced Auto-fluorescence: Auto-fluorescence is a common issue encountered in imaging biological samples and can interfere with the accurate detection of fluorescence signals. The developed optical imaging system employs appropriate filters and techniques to minimize autofluorescence, resulting in improved image quality and a better signal-to-noise ratio.
Integrated Image Capture and Analysis: The present invention integrates image capture and analysis functions, providing a streamlined workflow. The open-source Amcap software used in the system enables real-time image acquisition and analysis. By overcoming these drawbacks, the present invention offers a significant improvement in optical imaging technology, enabling researchers to conduct high-quality imaging studies at a reduced cost and with enhanced performance.

[07] IMPACT OF THE INVENTION
Fluorescence imaging systems have a significant impact across various fields, including biology, medicine, and materials science.
Biological Research: Fluorescence imaging allows researchers to visualize and track cellular processes in real-time, enabling insights into cell dynamics, protein interactions, and genetic expression.
Medical Diagnostics: In clinical settings, fluorescence imaging aids in detecting diseases such as cancer by highlighting abnormal cells or tissues, improving early diagnosis and treatment outcomes.
Drug Development: This technology is crucial in pharmacology, where it helps in studying drug interactions at the molecular level, leading to more effective therapies.
Environmental Monitoring: Fluorescence techniques are used to detect pollutants and monitor environmental changes, enhancing our understanding of ecological systems.
Materials Science: In this field, fluorescence imaging assists in characterizing materials, studying surface properties, and developing new materials with specific optical properties.

[08] MOTIVATION OF INVENTION
The evolution of fluorescence imaging technologies, making them essential tools in modern scientific research and clinical practice which are very expensive. The present invention is compact and cost effective.
, Claims:I/We claim
1. An in-vivo examination apparatus for examining a living organism comprising:
a mounting stand to hold the components in place;
a base metal to bear the weight of instrument;
an image capturing device;
an optic filter unit;
a slotted filter holder;
a mobile holder; and
an enclosure to mount the apparatus,
wherein the plurality of an image capturing device can do low light capturing with good resolution, and
wherein the plurality of an optic filter device has six number of slots in the different frequency ranges within limits, and
wherein the mobile holder position a mobile to produce ultra violet light along with lens and diffuser mechanism.
2. An imaging system as claimed in 1, capture high quality images of fluorescent samples at lower concentration;
an image sample can be from different size variations like carbon quantum dot level, phantom gel and microfluidic gel;
3. An optic system to be used as claimed in 2, has concave lenses, long-pass filters, low-light detectors, and integrated LED light sources of different colours.
4. The enclosure as claimed in 1, minimizes light interference, create controlled environment, reduce background noise and ensure accurate measurement.
5. An optic filter unit according to claim 1, wherein the first filter is of 400nm size and then limited to 650nm size in steps.

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