In Vivo Multiphoton Microscopy of Deep Brain Tissue

2004 ◽  
Vol 91 (4) ◽  
pp. 1908-1912 ◽  
Author(s):  
Michael J. Levene ◽  
Daniel A. Dombeck ◽  
Karl A. Kasischke ◽  
Raymond P. Molloy ◽  
Watt W. Webb
Keyword(s):  
Minimally Invasive ◽  
Intact Animal ◽  
Multiphoton Microscopy ◽  
Wild Type Mouse ◽  
Disease Diagnosis ◽  
Cortical Layer ◽  
Optical Biopsy ◽  
Proper Function ◽  
Subcellular Resolution

Although fluorescence microscopy has proven to be one of the most powerful tools in biology, its application to the intact animal has been limited to imaging several hundred micrometers below the surface. The rest of the animal has eluded investigation at the microscopic level without excising tissue or performing extensive surgery. However, the ability to image with subcellular resolution in the intact animal enables a contextual setting that may be critical for understanding proper function. Clinical applications such as disease diagnosis and optical biopsy may benefit from minimally invasive in vivo approaches. Gradient index (GRIN) lenses with needle-like dimensions can transfer high-quality images many centimeters from the object plane. Here, we show that multiphoton microscopy through GRIN lenses enables minimally invasive, subcellular resolution several millimeters in the anesthetized, intact animal, and we present in vivo images of cortical layer V and hippocampus in the anesthetized Thy1-YFP line H mouse. Microangiographies from deep capillaries and blood vessels containing fluorescein-dextran and quantum dot-labeled serum in wild-type mouse brain are also demonstrated.

scholarly journals 1700 nm optical coherence microscopy enables minimally invasive, label-free, in vivo optical biopsy deep in the mouse brain

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Jun Zhu ◽  
Hercules Rezende Freitas ◽  
Izumi Maezawa ◽  
Lee-way Jin ◽  
Vivek J. Srinivasan
Keyword(s):  
Minimally Invasive ◽  
Mouse Brain ◽  
Numerical Aperture ◽  
Optical Biopsy ◽  
Optical Coherence ◽  
Label Free ◽  
Optical Coherence Microscopy ◽  
Minimal Invasiveness ◽  
Cortical Regions

AbstractIn vivo, minimally invasive microscopy in deep cortical and sub-cortical regions of the mouse brain has been challenging. To address this challenge, we present an in vivo high numerical aperture optical coherence microscopy (OCM) approach that fully utilizes the water absorption window around 1700 nm, where ballistic attenuation in the brain is minimized. Key issues, including detector noise, excess light source noise, chromatic dispersion, and the resolution-speckle tradeoff, are analyzed and optimized. Imaging through a thinned-skull preparation that preserves intracranial space, we present volumetric imaging of cytoarchitecture and myeloarchitecture across the entire depth of the mouse neocortex, and some sub-cortical regions. In an Alzheimer’s disease model, we report that findings in superficial and deep cortical layers diverge, highlighting the importance of deep optical biopsy. Compared to other microscopic techniques, our 1700 nm OCM approach achieves a unique combination of intrinsic contrast, minimal invasiveness, and high resolution for deep brain imaging.

scholarly journals Gastroscopic Image Graph: Application to Noninvasive Multitarget Tracking under Gastroscopy

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Bin Wang ◽  
Weiling Hu ◽  
Jiquan Liu ◽  
Jianmin Si ◽  
Huilong Duan
Keyword(s):  
Disease Diagnosis ◽  
Optical Biopsy ◽  
Graph Search ◽  
Gastric Disease ◽  
Multitarget Tracking ◽  
Search Problem ◽  
Stomach Body ◽  
Attractive Candidate ◽  
The Mean

Gastroscopic examination is one of the most common methods for gastric disease diagnosis. In this paper, a multitarget tracking approach is proposed to assist endoscopists in identifying lesions under gastroscopy. This approach analyzes numerous preobserved gastroscopic images and constructs a gastroscopic image graph. In this way, the deformation registration between gastroscopic images is regarded as a graph search problem. During the procedure, the endoscopist marks suspicious lesions on the screen and the graph is utilized to locate and display the lesions in the appropriate frames based on the calculated registration model. Compared to traditional gastroscopic lesion surveillance methods (e.g., tattooing or probe-based optical biopsy), this approach is noninvasive and does not require additional instruments. In order to assess and quantify the performance, this approach was applied to stomach phantom data andin vivodata. The clinical experimental results demonstrated that the accuracy at angularis, antral, and stomach body was 6.3 ± 2.4 mm, 7.6 ± 3.1 mm, and 7.9 ± 1.6 mm, respectively. The mean accuracy was 7.31 mm, average targeting time was 56 ms, and thePvalue was 0.032, which makes it an attractive candidate for clinical practice. Furthermore, this approach provides a significant reference for endoscopic target tracking of other soft tissue organs.

scholarly journals Digital Image Analysis Tools Developed by the Indiana O’Brien Center

2021 ◽  
Vol 12 ◽  
Author(s):  
Kenneth W. Dunn
Keyword(s):  
Image Analysis ◽  
Cell Biology ◽  
Multiphoton Microscopy ◽  
Optical Sectioning ◽  
Biomedical Image ◽  
The Past ◽  
New Methods ◽  
Analysis Tools ◽  
Subcellular Resolution

The scale and complexity of images collected in biological microscopy have grown enormously over the past 30 years. The development and commercialization of multiphoton microscopy has promoted a renaissance of intravital microscopy, providing a window into cell biology in vivo. New methods of optical sectioning and tissue clearing now enable biologists to characterize entire organs at subcellular resolution. New methods of multiplexed imaging support simultaneous localization of forty or more probes at a time. Exploiting these exciting new techniques has increasingly required biomedical researchers to master procedures of image analysis that were once the specialized province of imaging experts. A primary goal of the Indiana O’Brien Center has been to develop robust and accessible image analysis tools for biomedical researchers. Here we describe biomedical image analysis software developed by the Indiana O’Brien Center over the past 25 years.

In Vivo Deep-Brain Structural and Hemodynamic Multiphoton Microscopy Enabled by Quantum Dots

Nano Letters ◽  
2019 ◽  
Vol 19 (8) ◽  
pp. 5260-5265 ◽  
Author(s):  
Hongji Liu ◽  
Xiangquan Deng ◽  
Shen Tong ◽  
Chen He ◽  
Hui Cheng ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Multiphoton Microscopy ◽  
Deep Brain

scholarly journals Ferromagnetic soft catheter robots for minimally invasive bioprinting

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Cheng Zhou ◽  
Youzhou Yang ◽  
Jiaxin Wang ◽  
Qingyang Wu ◽  
Zhuozhi Gu ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
The Body ◽  
Magnetic Actuation ◽  
Internal Organs ◽  
Planar Surfaces ◽  
Reinforced Polymer ◽  
Direct Fabrication ◽  
Digitally Controlled

AbstractIn vivo bioprinting has recently emerged as a direct fabrication technique to create artificial tissues and medical devices on target sites within the body, enabling advanced clinical strategies. However, existing in vivo bioprinting methods are often limited to applications near the skin or require open surgery for printing on internal organs. Here, we report a ferromagnetic soft catheter robot (FSCR) system capable of in situ computer-controlled bioprinting in a minimally invasive manner based on magnetic actuation. The FSCR is designed by dispersing ferromagnetic particles in a fiber-reinforced polymer matrix. This design results in stable ink extrusion and allows for printing various materials with different rheological properties and functionalities. A superimposed magnetic field drives the FSCR to achieve digitally controlled printing with high accuracy. We demonstrate printing multiple patterns on planar surfaces, and considering the non-planar surface of natural organs, we then develop an in situ printing strategy for curved surfaces and demonstrate minimally invasive in vivo bioprinting of hydrogels in a rat model. Our catheter robot will permit intelligent and minimally invasive bio-fabrication.

scholarly journals Effect of ethanol and cocaine on [11C]MPC-6827 uptake in SH-SY5Y cells

2021 ◽  
Author(s):  
Naresh Damuka ◽  
Miranda Orr ◽  
Paul W. Czoty ◽  
Jeffrey L. Weiner ◽  
Thomas J. Martin ◽  
...  
Keyword(s):  
Mechanism Of Action ◽  
Ex Vivo ◽  
Neuroblastoma Cells ◽  
Proper Function ◽  
Brain Functions ◽  
Biodistribution Studies ◽  
Positron Emission ◽  
Vitro Binding

AbstractMicrotubules (MTs) are structural units in the cytoskeleton. In brain cells they are responsible for axonal transport, information processing, and signaling mechanisms. Proper function of these processes is critical for healthy brain functions. Alcohol and substance use disorders (AUD/SUDs) affects the function and organization of MTs in the brain, making them a potential neuroimaging marker to study the resulting impairment of overall neurobehavioral and cognitive processes. Our lab reported the first brain-penetrant MT-tracking Positron Emission Tomography (PET) ligand [11C]MPC-6827 and demonstrated its in vivo utility in rodents and non-human primates. To further explore the in vivo imaging potential of [11C]MPC-6827, we need to investigate its mechanism of action. Here, we report preliminary in vitro binding results in SH-SY5Y neuroblastoma cells exposed to ethanol (EtOH) or cocaine in combination with multiple agents that alter MT stability. EtOH and cocaine treatments increased MT stability and decreased free tubulin monomers. Our initial cell-binding assay demonstrated that [11C]MPC-6827 may have high affinity to free/unbound tubulin units. Consistent with this mechanism of action, we observed lower [11C]MPC-6827 uptake in SH-SY5Y cells after EtOH and cocaine treatments (e.g., fewer free tubulin units). We are currently performing in vivo PET imaging and ex vivo biodistribution studies in rodent and nonhuman primate models of AUD and SUDs and Alzheimer's disease.

scholarly journals Effect of valproic acid on functional bleb morphology in a rabbit model of minimally invasive surgery

2021 ◽  
pp. bjophthalmol-2020-318691
Author(s):  
Zhu Li Yap ◽  
Li-Fong Seet ◽  
Stephanie WL Chu ◽  
Li Zhen Toh ◽  
Farah Ilyana Ibrahim ◽  
...  
Keyword(s):  
Valproic Acid ◽  
Cell Growth ◽  
Minimally Invasive ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Rabbit Model ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Label Free ◽  
Collagen Fibres

AbstractPurposeTo determine the effect of valproic acid (VPA) on bleb morphology and scar characteristics in a rabbit model of minimally invasive glaucoma surgery (MIGS).MethodsNine New Zealand white rabbits were subjected to MIGS with intraoperative implantation of the PreserFlo MicroShunt. Rabbits were then administered with subconjunctival injections of phosphate buffered saline (PBS) (n=4) or with VPA (n=5). Bleb morphology was examined by slit-lamp biomicroscopy and in vivo confocal microscopy. Postoperative day 28 tissues were examined by immunohistochemical evaluation and label-free multiphoton microscopy to visualise the collagen matrix, by terminal deoxynucleotidyl transferase dUTP nick-end labelling assay and immunofluorescent labelling for Ki67 expression to detect apoptosis and cell growth, and by real-time quantitative PCR to measure Col1a1, Fn, and Smad6 transcript expression.ResultsVPA-treated blebs were detectable on day 28, while the PBS-treated blebs were not detectable by day 14. VPA-treated blebs were diffuse, extended posteriorly with near normal conjunctival vascularity and featured a combination of reticular/blurred stromal pattern with evidence of relatively large stromal cysts. Instead of the deposition of thick, disorganised collagen fibres characteristic of the PBS bleb, the VPA bleb contained conspicuously thinner collagen fibres which were associated with similarly thinner fibronectin fibres. In corroboration, Col1a1 and Fn mRNA expression was reduced in the VPA blebs, while increased Smad6 expression implicated the disruption of the transforming growth factor beta pathway. Apoptosis and cell growth profiles appeared similar with both treatments.ConclusionsThe results support the application of VPA to enhance bleb morphology associated with good bleb function in MIGS with no apparent cytotoxicity.

scholarly journals Glycogen synthesis in the perfused liver of adrenalectomized rats

1976 ◽  
Vol 156 (3) ◽  
pp. 585-592 ◽  
Author(s):  
P D Whitton ◽  
D A Hems
Keyword(s):  
Intact Animal ◽  
Glycogen Synthesis ◽  
Hepatic Metabolism ◽  
Hepatic Glycogen ◽  
Perfused Liver ◽  
Short Term ◽  
Glycogen Accumulation ◽  
Glycogen Deposition ◽  
Adrenalectomized Rats

1. A total loss of capacity for net glycogen synthesis was observed in experiments with the perfused liver of starved adrenalectomized rats. 2. This lesion was corrected by insulin or cortisol in vivo (over 2-5h), but not by any agent tested in perfusion. 3. The activity of glycogen synthetase a, and its increase during perfusion, in the presence of glucose plus glucogenic substrates, were proportional to the rate of net glycogen accumulation. 4. This complete inherent loss of capacity for glycogen synthesis after adrenalectomy is greater than any defect in hepatic metabolism yet reported in this situation, and is not explicable by a decrease in the rate of gluconegenesis (which supports glycogen synthesis in the liver of starved rats). The short-term (2-5h) stimulatory effect of glucocorticoids in the intact animal, on hepatic glycogen deposition, may be mediated partly through insulin action, although neither insulin or cortisol appear to act directly on the liver to stimulate glycogen synthesis.

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