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Journal of Visualized Experiments : JoVE Jun 2018Microglia are brain phagocytes that participate in brain homeostasis and continuously survey their environment for dysfunction, injury, and disease. As the first...
Microglia are brain phagocytes that participate in brain homeostasis and continuously survey their environment for dysfunction, injury, and disease. As the first responders, microglia have important functions to mitigate neuron and glia dysfunction, and in this process, they undergo a broad range of morphologic changes. Microglia morphologies can be categorized descriptively or, alternatively, can be quantified as a continuous variable for parameters such as cell ramification, complexity, and shape. While methods for quantifying microglia are applied to single cells, few techniques apply to multiple microglia in an entire photomicrograph. The purpose of this method is to quantify multiple and single cells using readily available ImageJ protocols. This protocol is a summary of the steps and ImageJ plugins recommended to convert fluorescence and bright-field photomicrographs into representative binary and skeletonized images and to analyze them using software plugins AnalyzeSkeleton (2D/3D) and FracLac for morphology data collection. The outputs of these plugins summarize cell morphology in terms of process endpoints, junctions, and length as well as complexity, cell shape, and size descriptors. The skeleton analysis protocol described herein is well suited for a regional analysis of multiple microglia within an entire photomicrograph or region of interest (ROI) whereas FracLac provides a complementary individual cell analysis. Combined, the protocol provides an objective, sensitive, and comprehensive assessment tool that can be used to stratify between diverse microglia morphologies present in the healthy and injured brain.
Topics: Animals; Immunohistochemistry; Mice; Microglia; Photomicrography; Rats
PubMed: 29939190
DOI: 10.3791/57648 -
Experimental Eye Research Apr 2015The interphotoreceptor matrix (IPM) is a highly organized structure with interconnected domains surrounding cone and rod photoreceptor cells and extends throughout the... (Review)
Review
The interphotoreceptor matrix (IPM) is a highly organized structure with interconnected domains surrounding cone and rod photoreceptor cells and extends throughout the subretinal space. Based on known roles of the extracellular matrix in other tissues, the IPM is thought to have several prominent functions including serving as a receptor for growth factors, regulating retinoid transport, participating in cytoskeletal organization in surrounding cells, and regulation of oxygen and nutrient transport. In addition, a number of studies suggest that the IPM also may play a significant role in the etiology of retinal degenerative disorders. In this review, we describe the present knowledge concerning the structure and function of the IPM under physiological and pathological conditions.
Topics: Extracellular Matrix; Humans; Photomicrography; Photoreceptor Cells, Vertebrate
PubMed: 25819450
DOI: 10.1016/j.exer.2015.02.017 -
Archives of Pathology & Laboratory... Dec 2015The field of pathology is driven by microscopic images. Educational activities for trainees and practicing pathologists alike are conducted through exposure to images of... (Review)
Review
CONTEXT
The field of pathology is driven by microscopic images. Educational activities for trainees and practicing pathologists alike are conducted through exposure to images of a variety of pathologic entities in textbooks, publications, online tutorials, national and international conferences, and interdepartmental conferences. During the past century and a half, photographic technology has progressed from primitive and bulky, glass-lantern projector slides to static and/or whole slide digital-image formats that can now be transferred around the world in a matter of moments via the Internet.
OBJECTIVE
To provide a historic and technologic overview of the evolution of microscopic-image photographic tools and techniques.
DATA SOURCES
Primary historic methods of microscopic image capture were delineated through interviews conducted with senior staff members in the Emory University Department of Pathology. Searches for the historic image-capturing methods were conducted using the Google search engine. Google Scholar and PubMed databases were used to research methods of digital photography, whole slide scanning, and smart phone cameras for microscopic image capture in a pathology practice setting.
CONCLUSIONS
Although film-based cameras dominated for much of the time, the rise of digital cameras outside of pathology generated a shift toward digital-image capturing methods, including mounted digital cameras and whole slide digital-slide scanning. Digital image capture techniques have ushered in new applications for slide sharing and second-opinion consultations of unusual or difficult cases in pathology. With their recent surge in popularity, we suspect that smart phone cameras are poised to become a widespread, cost-effective method for pathology image acquisition.
Topics: Databases, Factual; History, 17th Century; History, 18th Century; History, 19th Century; History, 20th Century; History, 21st Century; Humans; Image Processing, Computer-Assisted; Internet; Photomicrography; Telepathology
PubMed: 25989285
DOI: 10.5858/arpa.2014-0315-RA -
Reviews in Neurological Diseases 2011Many studies have shown that hypertensive ocular funduscopic abnormalities are clearly related to stroke, even after controlling for blood pressure and other vascular... (Review)
Review
Many studies have shown that hypertensive ocular funduscopic abnormalities are clearly related to stroke, even after controlling for blood pressure and other vascular risk factors. Retinal abnormalities indicative of a breakdown of the blood-retina barrier confer a greater increase in risk for stroke than sclerotic retinal changes. Similar retinal changes also have a positive relationship to stroke mortality. In addition, hypertensive ocular fundus abnormalities are reported to be associated with an increased risk for cognitive impairment, cerebral atrophy, progression of magnetic resonance imaging-defined white matter lesions, and subclinical infarction. Recent advances in fundus photography allow for improved accuracy and consistency in interpretation of funduscopic lesions, and improve the feasibility of screening for these abnormalities in at-risk patient populations. Evaluating the ocular fundus for signs of hypertensive retinopathy, in combination with an assessment of the presence or absence of other known vascular risk factors, may allow clinicians to further individualize a risk profile for stroke to each individual patient, thus permitting more accurate risk stratification and, potentially, guiding treatment strategies.
Topics: Adult; Aged; Aged, 80 and over; Brain; Comorbidity; Diagnostic Techniques, Ophthalmological; Fundus Oculi; Humans; Hypertensive Retinopathy; Middle Aged; Photomicrography; Predictive Value of Tests; Retina; Retinal Vessels; Risk Assessment; Risk Factors; Stroke; Vascular Diseases
PubMed: 21769065
DOI: No ID Found -
British Heart Journal Sep 1965
Topics: Aged; Amyloidosis; Digitalis Glycosides; Heart Diseases; Heart Failure; Humans; Photomicrography
PubMed: 5829755
DOI: 10.1136/hrt.27.5.711 -
Journal of Burn Care & Research :... 2011A major potential goal of burn therapy is to limit progression of partial- to full-thickness burns. To better test therapies, the authors developed and validated a...
A major potential goal of burn therapy is to limit progression of partial- to full-thickness burns. To better test therapies, the authors developed and validated a vertical progression porcine burn model in which partial-thickness burns treated with an occlusive dressing convert to full-thickness burns that heal with scarring and wound contraction. Forty contact burns were created on the backs and flanks of two young swine using a 150 g aluminum bar preheated to 70°C, 80°C, or 90°C for 20 or 30 seconds. The necrotic epidermis was removed and the burns were covered with a polyurethane occlusive dressing. Burns were photographed at 1, 24, and 48 hours as well as at 7, 14, 21, and 28 days postinjury. Full-thickness biopsies were obtained at 1, 4, 24, and 48 hours as well as at 7 and 28 days. The primary outcomes were presence of deep contracted scars and wound area 28 days after injury. Secondary outcomes were depth of injury, reepithelialization, and depth of scars. Data were compared across burn conditions using analysis of variance and χ(2) tests. Eight replicate burns were created with the aluminum bar using the following temperature/contact-time combinations: 70/20, 70/30, 80/20, 80/30, and 90/20. The percentage of burns healing with contracted scars were 70/20, 0%; 70/30, 25%; 80/20, 50%; 80/30, 75%; and 90/20, 100% (P = .05). Wound areas at 28 days by injury conditions were 70/20, 8.1 cm(2); 70/30, 7.8 cm(2); 80/20, 6.6 cm(2); 80/30, 4.9 cm(2); and 90/20, 4.8 cm(2) (P = .007). Depth of injury judged by depth of endothelial damage for the 80/20 and 80/30 burns at 1 hour was 36% and 60% of the dermal thickness, respectively. The depth of injury to the endothelial cells 1 hour after injury was inversely correlated with the degree of scar area (Pearson's correlation r = -.71, P < .001). Exposure of porcine skin to an aluminum bar preheated to 80°C for 20 or 30 seconds results initially in a partial-thickness burn that when treated with an occlusive dressing progresses to a full-thickness injury and heals with significant scarring and wound contracture.
Topics: Animals; Bandages; Burns; Chi-Square Distribution; Cicatrix; Disease Models, Animal; Endothelium; Female; Photomicrography; Statistics as Topic; Swine
PubMed: 21841494
DOI: 10.1097/BCR.0b013e31822dc439 -
The Journal of Parasitology Aug 1988The "true" coccidia (phylum Apicomplexa, suborder Eimeriina) constitute a large and heterogeneous group of parasitic protozoa. Despite the large number of described... (Review)
Review
The "true" coccidia (phylum Apicomplexa, suborder Eimeriina) constitute a large and heterogeneous group of parasitic protozoa. Despite the large number of described species (ca. 1,650) and the medical and veterinary importance of some (e.g., Toxoplasma), 2 facts are clear: (1) the majority of coccidia species are probably yet undescribed, and (2) the phylogenetic relationships of those described species are poorly known. Contributing to the latter dilemma is the lack of a tradition to provide type specimens by those who describe new species, even though the International Code of Zoological Nomenclature specifically recommends the designation of a type specimen with the description of a new species. With the publication of a new edition of the Code (1985), explicit provisions are made for the unique concerns of taxonomists working with Protozoa. Here we remind those interested in the taxonomy of coccidia of an already established method for preserving oocysts in resin and, as an alternative, suggest the standardization of a photographic procedure through which type specimens of coccidia oocysts might also be submitted to and maintained in accredited museums. Thus, coccidia taxonomists should no longer have an excuse for their failure to designate types.
Topics: Animals; Coccidia; Photomicrography; Preservation, Biological; Terminology as Topic
PubMed: 3294365
DOI: No ID Found -
Journal of Clinical Laboratory Analysis 2004The computer and the digital camera provide a unique means for improving hematology education, research, and patient service. High quality photographic images of gross... (Review)
Review
The computer and the digital camera provide a unique means for improving hematology education, research, and patient service. High quality photographic images of gross specimens can be rapidly and conveniently acquired with a high-resolution digital camera, and specialized digital cameras have been developed for photomicroscopy. Digital cameras utilize charge-coupled devices (CCD) or Complementary Metal Oxide Semiconductor (CMOS) image sensors to measure light energy and additional circuitry to convert the measured information into a digital signal. Since digital cameras do not utilize photographic film, images are immediately available for incorporation into web sites or digital publications, printing, transfer to other individuals by email, or other applications. Several excellent digital still cameras are now available for less than 2,500 dollars that capture high quality images comprised of more than 6 megapixels. These images are essentially indistinguishable from conventional film images when viewed on a quality color monitor or printed on a quality color or black and white printer at sizes up to 11x14 inches. Several recent dedicated digital photomicroscopy cameras provide an ultrahigh quality image output of more than 12 megapixels and have low noise circuit designs permitting the direct capture of darkfield and fluorescence images. There are many applications of digital images of pathologic specimens. Since pathology is a visual science, the inclusion of quality digital images into lectures, teaching handouts, and electronic documents is essential. A few institutions have gone beyond the basic application of digital images to developing large electronic hematology atlases, animated, audio-enhanced learning experiences, multidisciplinary Internet conferences, and other innovative applications. Digital images of single microscopic fields (single frame images) are the most widely utilized in hematology education at this time, but single images of many adjacent microscopic fields can be stitched together to prepare "zoomable" panoramas that encompass a large part of a microscope slide and closely simulate observation through a real microscope. With further advances in computer speed and Internet streaming technology, the virtual microscope could easily replace the real microscope in pathology education. Later in this decade, interactive immersive computer experiences may completely revolutionize hematology education and make the conventional lecture and laboratory format obsolete. Patient care is enhanced by the transmission of digital images to other individuals for consultation and education, and by the inclusion of these images in patient care documents. In research laboratories, digital cameras are widely used to document experimental results and to obtain experimental data.
Topics: Computers; Humans; Image Processing, Computer-Assisted; Information Storage and Retrieval; Microscopy, Video; Pathology, Clinical; Photography; Photomicrography; Quality Assurance, Health Care; Software; Telepathology
PubMed: 15065212
DOI: 10.1002/jcla.20009 -
The Journal of Urology Oct 2008Although genital tactile stimulation is regarded as a precursor to sexual arousal and a recognized initiator of central nervous system arousal, specific afferent neural... (Review)
Review
PURPOSE
Although genital tactile stimulation is regarded as a precursor to sexual arousal and a recognized initiator of central nervous system arousal, specific afferent neural pathways transmit sensory stimuli of arousal, beginning at the epithelial level on the clitoris and following the course of arousal stimuli through the central nervous system. Limited knowledge exists of the pathway from the cutaneous receptors of nerves originating in the epithelial tissue of the clitoris and continuing to spinal cord afferents. Such information may contribute to an understanding of sexual arousal, particularly in female vertebrates. We further defined the neural pathways and mechanisms responsible for arousal originating in the epithelium of the clitoris as well as related neural pathways to the spinal cord in a murine model.
MATERIALS AND METHODS
We performed a comprehensive review of the published relevant clinical and histological material from human and nonhuman vertebrate studies. In 29 adult female C57B1/6 mice the distribution of pelvic nerves and vessels was mapped. Gross dissection of 4 female mice was facilitated by resin injection of the vascular system in 2. Neuronal tracing was performed in 25 mice that received clitoral injection of wheat germ agglutinin-horseradish peroxidase into the clitoris and were sacrificed after 72 to 96 hours. The spinal cord and periclitoral tissue were removed and fixed. Immunohistochemistry was performed.
RESULTS
Gross anatomy of the mouse clitoris showed that pudendal and hypogastric nerves have a major role in the innervation of the external genitalia. Neuronal tracing revealed that the greatest nerve density was noted in the L5/6 spinal cord. The distribution extended from S1 to L2 with no labeling seen in the L3 spinal cord. Wheat germ agglutinin-horseradish peroxidase labeling was seen caudal in levels S1 through L4 and rostral in L2.
CONCLUSIONS
Understanding the neuroanatomy of the clitoris using a murine model may provide a valuable tool for the study of sexual arousal disorders and the further understanding of sexual function related to neural pathologies and trauma.
Topics: Afferent Pathways; Animals; Arousal; Clitoris; Dissection; Female; Horseradish Peroxidase; Mice; Mice, Inbred BALB C; Models, Animal; Neurons, Afferent; Orgasm; Photomicrography; Sensitivity and Specificity; Sexual Behavior, Animal; Spinal Cord; Wheat Germ Agglutinins
PubMed: 18707740
DOI: 10.1016/j.juro.2008.06.009 -
Imaging of molecular dynamics regulated by electrical activities in neural circuits and in synapses.Neuro-Signals 2008One of the major challenges in brain research is to unravel a network of molecules, neurons, circuits and systems that are responsible for dynamic and hierarchical brain... (Review)
Review
One of the major challenges in brain research is to unravel a network of molecules, neurons, circuits and systems that are responsible for dynamic and hierarchical brain functions. To understand molecular events that occur in synapses could be an important key to exploring the mechanism of information processing. A spatiotemporal recording method is required to observe neuronal activities in a particular local circuit and to resolve single synaptic potential with high resolution. As alternative methods, real-time imaging using fluorescent probes and optical recording methods are also a powerful approach for investigating the molecular dynamics of biological events in neurons in vitro and in vivo. Recently, optical imaging techniques have become of great importance to visualize the molecular dynamics in a micron-sized compartment of a single neuron such as neuronal synapse. In general, the presynaptic axon forms synapses at the postsynaptic site on the dendritic spines in the mammalian central nervous system. Subsets of the synapses undergo a series of enduring changes in spine shape and density as well as alterations in electrophysiological functions. Here we describe recent optical imaging studies conducted by elaborate methods and techniques that provide evidence for the link between neural activity and molecular dynamics.
Topics: Animals; Biological Transport; Brain; Cells, Cultured; Computer Systems; Dendrites; Fiber Optic Technology; Fluorescent Dyes; Forecasting; Hippocampus; Humans; Microscopy, Fluorescence; Nerve Net; Nerve Tissue Proteins; Neuroglia; Neurons; Patch-Clamp Techniques; Photomicrography; Pyridinium Compounds; RNA, Messenger; RNA-Binding Proteins; Synaptic Transmission
PubMed: 18635943
DOI: 10.1159/000123037