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Time-Lapse Visualization of Microglia Cell Processes using Fluorescent Miniature (Miniscope) Imaging

Description

In the United States, an estimated 2 million cases of traumatic brain injury (TBI) resulting in more than 50,000 deaths occur every year. TBI induces an immediate primary injury resulting in local or diffuse cell death in the brain. Then

In the United States, an estimated 2 million cases of traumatic brain injury (TBI) resulting in more than 50,000 deaths occur every year. TBI induces an immediate primary injury resulting in local or diffuse cell death in the brain. Then a secondary injury occurs through neuroinflammation from immune cells in response to primary injury. Microglia, the resident immune cell of the central nervous system, play a critical role in neuroinflammation following TBI. Microglia make up 10% of all cells in the nervous system and are the fastest moving cells in the brain, scanning the entire parenchyma every several hours. Microglia have roles in both the healthy and injured brain. In the healthy brain, microglia can produce neuroprotective factors, clear cellular debris, and organize neurorestorative processes to recover from TBI. However, microglia mediated neuroinflammation during secondary injury produces pro-inflammatory and cytotoxic mediators contributing to neuronal dysfunction, inhibition of CNS repair, and cell death. Furthermore, neuroinflammation is a prominent feature in many neurodegenerative diseases such as Alzheimer’s, and Parkinson’s disease, of which include overactive microglia function. Microglia cell morphology, activation, and response to TBI is poorly understood. Currently, imaging microglia can only be performed while the animal is stationary and under anesthesia. The Miniscope technology allows for real-time visualization of microglia in awake behaving animals. The Miniscope is a miniature fluorescent microscope that can be implanted over a craniectomy to image microglia. Currently, the goals of Miniscope imaging are to improve image quality and develop time-lapse imaging capabilities. There were five main sub-projects that focused on these goals including surgical nose cone design, surgical holder design, improved GRIN lens setup, improved magnification through achromatic lenses, and time-lapse imaging hardware development. Completing these goals would allow for the visualization of microglia function in the healthy and injured brain, elucidating important immune functions that could provide new strategies for treating brain diseases.

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2019-05

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Image Analysis for Registration, Segmentation, and Intensity Measurement of 2-Photon Microscope Images using MATLAB

Description

Traumatic brain injury (TBI) is a major concern in public health due to its prevalence and effect. Every year, about 1.7 million TBIs are reported [7]. According to the According to the Centers for Disease Control and Prevention (CDC), 5.5%

Traumatic brain injury (TBI) is a major concern in public health due to its prevalence and effect. Every year, about 1.7 million TBIs are reported [7]. According to the According to the Centers for Disease Control and Prevention (CDC), 5.5% of all emergency department visits, hospitalizations, and deaths from 2002 to 2006 are due to TBI [8]. The brain's natural defense, the Blood Brain Barrier (BBB), prevents the entry of most substances into the brain through the blood stream, including medicines administered to treat TBI [11]. TBI may cause the breakdown of the BBB, and may result in increased permeability, providing an opportunity for NPs to enter the brain [3,4]. Dr. Stabenfeldt's lab has previously established that intravenously injected nanoparticles (NP) will accumulate near the injury site after focal brain injury [4]. The current project focuses on confirmation of the accumulation or extravasation of NPs after brain injury using 2-photon microscopy. Specifically, the project used controlled cortical impact injury induced mice models that were intravenously injected with 40nm NPs post-injury. The MATLAB code seeks to analyze the brain images through registration, segmentation, and intensity measurement and evaluate if fluorescent NPs will accumulate in the extravascular tissue of injured mice models. The code was developed with 2D bicubic interpolation, subpixel image registration, drawn dimension segmentation and fixed dimension segmentation, and dynamic image analysis. A statistical difference was found between the extravascular tissue of injured and uninjured mouse models. This statistical difference proves that the NPs do extravasate through the permeable cranial blood vessels in injured cranial tissue.

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2018-05

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Developing a Virtual Heart Library for Use in Pediatric Heart Transplant Allograft Size Selection

Description

Introduction: There are 350 to 400 pediatric heart transplants annually according to the Pediatric Heart Transplant Database (Dipchand et al. 2014). Finding appropriate donors can be challenging especially for the pediatric population. The current standard of care

Introduction: There are 350 to 400 pediatric heart transplants annually according to the Pediatric Heart Transplant Database (Dipchand et al. 2014). Finding appropriate donors can be challenging especially for the pediatric population. The current standard of care is a donor-to-recipient weight ratio. This ratio is not necessarily a parameter directly indicative of the size of a heart, potentially leading to ill-fitting allografts (Tang et al. 2010). In this paper, a regression model is presented - developed by correlating total cardiac volume to non-invasive imaging parameters and patient characteristics – for use in determining ideal allograft fit with respect to total cardiac volume.
Methods: A virtual, 3D library of clinically-defined normal hearts was compiled from reconstructed CT and MR scans. Non-invasive imaging parameters and patient characteristics were collected and subjected to backward elimination linear regression to define a model relating patient parameters to the total cardiac volume. This regression model was then used to retrospectively accept or reject an ‘ideal’ donor graft from the library for 3 patients that had undergone heart transplantation. Oversized and undersized grafts were also transplanted to qualitatively analyze virtual transplantation specificity.
Results: The backward elimination approach of the data for the 20 patients rejected the factors of BMI, BSA, sex and both end-systolic and end-diastolic left ventricular measurements from echocardiography. Height and weight were included in the linear regression model yielding an adjusted R-squared of 82.5%. Height and weight showed statistical significance with p-values of 0.005 and 0.02 respectively. The final equation for the linear regression model was TCV = -169.320+ 2.874h + 3.578w ± 73 (h=height, w=weight, TCV= total cardiac volume).
Discussion: With the current regression model, height and weight significantly correlate to total cardiac volume. This regression model and virtual normal heart library provide for the possibility of virtual transplant and size-matching for transplantation. The study and regression model is, however, limited due to a small sample size. Additionally, the lack of volumetric resolution from the MR datasets is a potentially limiting factor. Despite these limitations the virtual library has the potential to be a critical tool for clinical care that will continue to grow as normal hearts are added to the virtual library.

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Date Created
2016-05

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Electromyograph Remote Control Jellyfish Toy: A Brief Exploration of Jellyfish Biomimetics

Description

The goal of this project was to explore biomimetics by creating a jellyfish flying device that uses propulsion of air to levitate while utilizing electromyography signals and infrared signals as mechanisms to control the device. Completing this project would require

The goal of this project was to explore biomimetics by creating a jellyfish flying device that uses propulsion of air to levitate while utilizing electromyography signals and infrared signals as mechanisms to control the device. Completing this project would require knowledge of biological signals, electrical circuits, computer programming, and physics to accomplish. An EMG sensor was used to obtain processed electrical signals produced from the muscles in the forearm and was then utilized to control the actuation speed of the tentacles. An Arduino microprocessor was used to translate the EMG signals to infrared blinking sequences which would propagate commands through a constructed circuit shield to the infrared receiver on jellyfish. The receiver will then translate the received IR sequence into actions. Then the flying device must produce enough thrust to propel the body upwards. The application of biomimetics would best test my skills as an engineer as well as provide a method of applying what I have learned over the duration of my undergraduate career.

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Date Created
2014-05

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Novel siloxane nanoprobes for molecular and cellular imagin

Description

Oxygen delivery is crucial for the development of healthy, functional tissue. Low tissue oxygenation, or hypoxia, is a characteristic that is common in many tumors. Hypoxia contributes to tumor malignancy and can reduce the success of chemotherapy and radiation treatment.

Oxygen delivery is crucial for the development of healthy, functional tissue. Low tissue oxygenation, or hypoxia, is a characteristic that is common in many tumors. Hypoxia contributes to tumor malignancy and can reduce the success of chemotherapy and radiation treatment. There is a current need to noninvasively measure tumor oxygenation or pO2 in patients to determine a personalized treatment method. This project focuses on creating and characterizing nanoemulsions using a pO2 reporter molecule hexamethyldisiloxane (HMDSO) and its longer chain variants as well as assessing their cytotoxicity. We also explored creating multi-modal (MRI/Fluorescence) nanoemulsions.

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Date Created
2013-05

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A Low Cost, High Dynamic Range, Versatile Digital Readout Integrated Circuit Unit Cell Prototype for Infrared Imaging Applications

Description

Readout Integrated Circuits(ROICs) are important components of infrared(IR) imag

ing systems. Performance of ROICs affect the quality of images obtained from IR

imaging systems. Contemporary infrared imaging applications demand ROICs that

can support large dynamic range, high frame rate, high output data rate,

Readout Integrated Circuits(ROICs) are important components of infrared(IR) imag

ing systems. Performance of ROICs affect the quality of images obtained from IR

imaging systems. Contemporary infrared imaging applications demand ROICs that

can support large dynamic range, high frame rate, high output data rate, at low

cost, size and power. Some of these applications are military surveillance, remote

sensing in space and earth science missions and medical diagnosis. This work focuses

on developing a ROIC unit cell prototype for National Aeronautics and Space Ad

ministration(NASA), Jet Propulsion Laboratory’s(JPL’s) space applications. These

space applications also demand high sensitivity, longer integration times(large well

capacity), wide operating temperature range, wide input current range and immunity

to radiation events such as Single Event Latchup(SEL).

This work proposes a digital ROIC(DROIC) unit cell prototype of 30ux30u size,

to be used mainly with NASA JPL’s High Operating Temperature Barrier Infrared

Detectors(HOT BIRDs). Current state of the art DROICs achieve a dynamic range

of 16 bits using advanced 65-90nm CMOS processes which adds a lot of cost overhead.

The DROIC pixel proposed in this work uses a low cost 180nm CMOS process and

supports a dynamic range of 20 bits operating at a low frame rate of 100 frames per

second(fps), and a dynamic range of 12 bits operating at a high frame rate of 5kfps.

The total electron well capacity of this DROIC pixel is 1.27 billion electrons, enabling

integration times as long as 10ms, to achieve better dynamic range. The DROIC unit

cell uses an in-pixel 12-bit coarse ADC and an external 8-bit DAC based fine ADC.

The proposed DROIC uses layout techniques that make it immune to radiation up to

300krad(Si) of total ionizing dose(TID) and single event latch-up(SEL). It also has a

wide input current range from 10pA to 1uA and supports detectors operating from

Short-wave infrared (SWIR) to longwave infrared (LWIR) regions.

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Date Created
2019