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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 -
Journal of Microscopy and Ultrastructure 2022Smartphones can be used to capture images from the microscope. There are commercial and homemade adapters that can be used to overcome the difficulty of focusing on a...
BACKGROUND AND AIM
Smartphones can be used to capture images from the microscope. There are commercial and homemade adapters that can be used to overcome the difficulty of focusing on a smartphone camera. We conducted this study to test if the usage of a homemade adapter reduces the time and effort of the operator in comparison to the free-hand technique in smartphone photomicrography.
MATERIALS AND METHODS
We made a simple smartphone adapter for digital photomicrography. Thirty-two operators first captured the image on the smartphone with the free-hand technique and then with the adapter thrice from a microscope. The time for focusing was compared statistically by paired -test. A survey was conducted to know operators's opinions on adapter-assisted photomicrography.
RESULT
All the participants were able to capture images from the microscopes. The average time for focusing with adapter was (11.89 ± 4.19 seconds) significantly ( = 0.0001) lower than the free-hand technique (25.56±11.81 seconds). However, the images yielded from both techniques were of equal quality. The majority of the participant found the method to be easy and reduce their effort in focusing and capturing an image.
CONCLUSION
Low-cost homemade smartphone adapter helps in reducing the time required for capturing an image from the microscope. It decreases the effort of the operator in comparison to the free-hand technique. Hence, it may be a good choice for those who need to capture images from the microscopes frequently for either telemedicine or research purpose in resource-limited settings.
PubMed: 35433256
DOI: 10.4103/JMAU.JMAU_45_20 -
Journal of Family Medicine and Primary... May 2020In recent years, a new concept has emerged at the forefront of slide-based diagnosis and telepathology. This is the concept of whole slide imaging. This has very...
CONTEXT
In recent years, a new concept has emerged at the forefront of slide-based diagnosis and telepathology. This is the concept of whole slide imaging. This has very recently also been tried with smartphones using complex software for photo analysis and stitching. This study is aimed to evaluate and compare the use of Jenoptik photomicrograph camera and smartphone camera-based whole slide imaging (WSI) for various microscopic preparations and slides. The study also uses a commonly available IOS smartphone device, a commonly available phone to microscope attachment, and the most commonly used Adobe Photoshop software for all stitching purposes.
AIM
To evaluate and compare smartphone and photomicrography-based whole slide imaging.
SUBJECTS AND METHODS
The study was conducted at the Department of Oral Pathology and Microbiology in Bapuji Dental College and Hospital, Davangere. The samples included in this study are 10 slides of routine hematoxylin and eosin stain slides and 10 ground section slides of teeth and bone samples. Continuous photographs of the entire slide were captured using an IOS device and a Jenoptik camera attached research microscope at 10× magnification. The photographs were stitched using Adobe Photoshop 2017 software to obtain a whole slide image. Then, two observers analyzed the whole slide image for a possible diagnosis and thus compared the efficiency of both the methods.
STATISTICAL ANALYSIS USED
The statistical Chi-square test and analysis of variance (ANOVA) H test were done using Statistical Package for the Social Sciences (SPSS) 2010 Software.
RESULTS
Percentage of measure of agreement was 79.5%, 83.7% and 86.3%, 89.1% for the whole slide images taken in the smartphone and photomicrograph and analyzed by Observer 1 and Observer 2, respectively.
CONCLUSIONS
In this study, it can be seen that the WSI by research microscopes is better than that with a smartphone and a compound microscope but still both the methods are equally good and can be followed with accurate results.
PubMed: 32754495
DOI: 10.4103/jfmpc.jfmpc_18_20 -
Microorganisms Jan 2019Dinoflagellates have some of the largest genome sizes, but lack architectural nucleosomes. Their liquid crystalline chromosomes (LCCs) are the only non-architectural... (Review)
Review
Dinoflagellates have some of the largest genome sizes, but lack architectural nucleosomes. Their liquid crystalline chromosomes (LCCs) are the only non-architectural protein-mediated chromosome packaging systems, having high degrees of DNA superhelicity, liquid crystalline condensation and high levels of chromosomal divalent cations. Recent observations on the reversible decompaction⁻recompaction of higher-order structures implicated that LCCs are composed of superhelical modules (SPMs) comprising highly supercoiled DNA. Orientated polarizing light photomicrography suggested the presence of three compartments with different packaging DNA density in LCCs. Recent and previous biophysical data suggest that LCCs are composed of: (a) the highly birefringent inner core compartment (i) with a high-density columnar-hexagonal mesophase (CH-m); (b) the lower-density core surface compartment (ii.1) consisting of a spiraling chromonema; (c) the birefringent-negative periphery compartment (ii.2) comprising peripheral chromosomal loops. C(ii.1) and C(ii.2) are in dynamic equilibrium, and can merge into a single compartment during dinomitosis, regulated through multiphasic reversible soft-matter phase transitions.
PubMed: 30678153
DOI: 10.3390/microorganisms7020027 -
Frontiers in Plant Science 2023Oil-based emulsion solution is a common pesticide formulation in agricultural spraying, and its spray characteristics are different from that of water spraying. The well...
INTRODUCTION
Oil-based emulsion solution is a common pesticide formulation in agricultural spraying, and its spray characteristics are different from that of water spraying. The well understanding of its spray characteristics is the theoretical basis to improve the pesticide spraying technology. The objective of the present study is to deepen the understanding of the spray characteristics of oil-based emulsion.
METHOD
In this paper, the spatial distribution characteristics of spray droplets of oil-based emulsion were captured visually using the high-speed photomicrography. On the basis of image processing method, the droplet size and distribution density of spray droplets at different spatial locations were analyzed quantitatively. The effects of nozzle configuration and emulsion concentration on spray structures and droplet spatial distribution were discussed.
RESULTS
Oil-based emulsion produced a special perforation atomization mechanism compared with water spray, which led to the increase of spray droplet size and distribution density. Nozzle configuration had a significant effect on oil-based emulsion spray, with the nozzle changed from ST110-01 to ST110-03 and ST110-05; the sheet lengths increased to 18 and 28 mm, respectively, whereas the volumetric median diameters increased to 51.19% and 76.00%, respectively. With emulsion concentration increased from 0.02% to 0.1% and 0.5%, the volumetric median diameters increased to 5.17% and 14.56%, respectively.
DISCUSSION
The spray droplet size of oil-based emulsion spray can be scaled by the equivalent diameter of discharge orifice of nozzles. The products of volumetric median diameters and corresponding surface tensions were nearly constant for the oil-based emulsion spray of different emulsion concentrations. It is expected that this research could provide theoretical support for improving the spraying technology of oil-based emulsion and increasing the utilization of pesticide.
PubMed: 37360725
DOI: 10.3389/fpls.2023.1183387 -
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 -
Journal of Microscopy and Ultrastructure 2020Modern light microscopes are available with built-in illuminator and facility of photomicrography. This enables the microscopy to be ready for telemedicine. However,...
INTRODUCTION
Modern light microscopes are available with built-in illuminator and facility of photomicrography. This enables the microscopy to be ready for telemedicine. However, resource-limited settings still find difficulty in procuring those microscopes.
AIM
The aim of this study was to upgrade a light microscope to a smartphone-connected digital microscope with minimal cost to make it ready for telemedicine.
MATERIALS AND METHODS
A commercially available (price: ₹389) Universal Serial Bus (USB) web camera was set on the eyepiece and fixed with the help of an aluminum sheet. Light emitting diodes (LEDs), covered with an optical diffuser, were set below the condenser. The camera was connected to an Android smartphone with an application for capturing image and video. Sixteen faculty members provided their opinion about the new device.
RESULTS
The smartphone-connected microscope was successfully used to focus and capture image and video of various slides. The images and videos were stored in the smartphone and shared via E-mail and other channels (e.g., WhatsApp and Telegram). This camera was also successfully connected to a laptop for projecting the real-time microscopic field on a screen. According to faculty members, focusing an object and capturing the image are the best features of the device; however, development of the device received lowest score.
CONCLUSION
A light microscope was upgraded to telemedicine ready microscope with nominal cost and moderate effort. It can also be used in medical teachings as it can project real-time images of a slide under the microscope. As it is equipped with LEDs, powered by the same smartphone, it can be operated without daylight or during a power outage.
PubMed: 32766118
DOI: 10.4103/JMAU.JMAU_35_19 -
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 -
Investigative Ophthalmology & Visual... Dec 2019The subbasal nerve plexus (SNP) is the densest and most recognizable component of the mammalian corneal innervation; however, the anatomical configuration of the SNP in...
PURPOSE
The subbasal nerve plexus (SNP) is the densest and most recognizable component of the mammalian corneal innervation; however, the anatomical configuration of the SNP in most animal models remains incompletely described. The purpose of the current study is to describe in detail the SNP architecture in eight different mammals, including several popular animal models used in cornea research.
METHODS
Corneal nerves in mouse, rat, guinea pig, rabbit, dog, macaque, domestic pig, and cow eyes were stained immunohistochemically with antiserum directed against neurotubulin. SNP architecture was documented by digital photomicrography and large-scale reconstructions, that is, corneal nerve maps, using a drawing tube attached to a light microscope.
RESULTS
Subbasal nerve fibers (SNFs) in mice, rats, guinea pigs, dogs, and macaques radiated centrally from the corneoscleral limbus toward the corneal apex in a whorl-like or spiraling pattern. SNFs in rabbit and bovine corneas swept horizontally across the ocular surface in a temporal-to-nasal direction and converged on the inferonasal limbus without forming a spiral. SNFs in the pig cornea radiated centrifugally in all directions, like a starburst, from a focal point located equidistant between the corneal apex and the superior pole.
CONCLUSIONS
The results of the present study have demonstrated for the first time substantial interspecies differences in the architectural organization of the mammalian SNP. The physiological significance of these different patterns and the mechanisms that regulate SNP pattern formation in the mammalian cornea remain incompletely understood and warrant additional investigation.
Topics: Anatomy, Comparative; Animals; Cattle; Cornea; Dogs; Guinea Pigs; Macaca; Mice; Microscopy, Confocal; Models, Animal; Nerve Fibers; Ophthalmic Nerve; Rabbits; Rats; Swine; Trigeminal Ganglion
PubMed: 31790560
DOI: 10.1167/iovs.19-28519 -
Neuroscience Mar 2017Cortico-cortical connectivity has become a major focus of neuroscience in the last decade but most of the connectivity studies focused on intrahemispheric circuits....
Cortico-cortical connectivity has become a major focus of neuroscience in the last decade but most of the connectivity studies focused on intrahemispheric circuits. Little has been reported about information acquired and processed in the premotor cortex and its functional connection with its homotopic counterpart in the opposite hemisphere via the corpus callosum. In non-human primates (macaques) lateralization is not well documented and its exact role is still unknown. The present study confirms in two macaques the existence of homotopic contralateral projections and completes the picture by further exploring heterotopic (non-motor) callosal projections. This was tested by injecting retrograde tracers in the premotor cortical areas PMv and PMd (targets). Our method consisted of identifying the connections with all the homo- and heterotopic cortical areas located in the contralateral hemisphere. The results showed that PMd and PMv receive multiple low-density labeled inputs from the opposite heterotopic prefrontal, parietal, motor, insular and temporal regions. Such unexpected collection of transcallosal inputs from heterotopic areas suggests that the premotor areas communicate with other modalities through long distance low-density networks which could have important implications in the understanding of sensorimotor and multimodal integration.
Topics: Animals; Corpus Callosum; Functional Laterality; Macaca fascicularis; Macaca mulatta; Motor Cortex; Neural Pathways; Neuroanatomical Tract-Tracing Techniques; Neurons; Photomicrography
PubMed: 28042027
DOI: 10.1016/j.neuroscience.2016.12.035