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Bio-medical Materials and Engineering 2020Surface replication is a nondestructive evaluation technique applied in examining surface wear by recording surface irregularities, especially in conditions when...
BACKGROUND
Surface replication is a nondestructive evaluation technique applied in examining surface wear by recording surface irregularities, especially in conditions when surfaces of interest cannot be further manipulated to fit directly under a microscope to be examined. Enamel is the outermost protective layer of the human teeth and is constantly stressed by mastication forces which results in enamel wear.
OBJECTIVE
To date, a procedure combining the clinical and microscopic examination of enamel surfaces is absent, which hinders the early diagnosis and comprehension of the wear process.
METHODS
This study investigated the role of replication sheets in registering microscopic wear on human enamel surfaces by both negative and positive replication techniques.
RESULTS
The sheets replicated wear features successfully. Sheets were compatible to use with multiple microscopes, with proper preparation, including high resolution microscopes such as the scanning electron microscope and transmitting electron microscope.
Topics: Dental Enamel; Humans; Microscopy, Electron, Scanning; Surface Properties
PubMed: 32986650
DOI: 10.3233/BME-206006 -
Current Medical Imaging Jun 2023Deconvolution microscopy is a computational image-processing technique used in conjunction with fluorescence microscopy to increase the resolution and contrast of...
Deconvolution microscopy is a computational image-processing technique used in conjunction with fluorescence microscopy to increase the resolution and contrast of three-dimensional images. Fluorescence microscopy is a widely used technique in biology and medicine that involves labeling specific molecules or structures within a sample with fluorescent dyes and then electronically photographing the sample through a microscope. However, the resolution of conventional fluorescence microscopy is limited by diffraction within the microscope's optical path, which causes blurring of the image and reduces the ability to resolve structures in close proximity with one another. Deconvolution microscopy overcomes this limitation by means of computer-based image processing whereby mathematical algorithms are used to eliminate the blurring caused by the microscope's optics and thus obtain a higher-resolution image that reveals the fine details of the sample with greater accuracy. Deconvolution microscopy, which can be applied to a range of image acquisition modalities, including widefield, confocal, and super-resolution microscopy, has become an essential tool for studying the structure and function of biological systems at the cellular and molecular levels. In this perspective, the latest deconvolution techniques have been introduced and image-processing methods for medical purposes have been presented.
PubMed: 37272458
DOI: 10.2174/1573405620666230602123028 -
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 -
Biophysical Reports Sep 2021One of the most widely used microscopy techniques in biology and medicine is fluorescence microscopy, offering high specificity in labeling as well as maximal... (Review)
Review
One of the most widely used microscopy techniques in biology and medicine is fluorescence microscopy, offering high specificity in labeling as well as maximal sensitivity. For live-cell imaging, the ideal fluorescence microscope should offer high spatial resolution, fast image acquisition, three-dimensional sectioning, and multicolor detection. However, most existing fluorescence microscopes have to compromise between these different requirements. Here, we present a multiplane, multicolor wide-field microscope that uses a dedicated beam splitter for recording volumetric data in eight focal planes and for three emission colors with frame rates of hundreds of volumes per second. We demonstrate the efficiency and performance of our system by three-dimensional imaging of multiply labeled fixed and living cells. The use of commercially available components makes our proposed microscope straightforward for implementation, thus promising for widely used applications.
PubMed: 36425311
DOI: 10.1016/j.bpr.2021.100001 -
Journal of Microscopy Mar 2021Correlative light and electron microscopy (CLEM) is a method used to investigate the exact same region in both light and electron microscopy (EM) in order to add...
Correlative light and electron microscopy (CLEM) is a method used to investigate the exact same region in both light and electron microscopy (EM) in order to add ultrastructural information to a light microscopic (usually fluorescent) signal. Workflows combining optical or fluorescent data with electron microscopic images are complex, hence there is a need to communicate detailed protocols and share tips & tricks for successful application of these methods. With the development of volume-EM techniques such as serial blockface scanning electron microscopy (SBF-SEM) and Focussed Ion Beam-SEM, correlation in three dimensions has become more efficient. Volume electron microscopy allows automated acquisition of serial section imaging data that can be reconstructed in three dimensions (3D) to provide a detailed, geometrically accurate view of cellular ultrastructure. In addition, combining volume-EM with high-resolution light microscopy (LM) techniques decreases the resolution gap between LM and EM, making retracing of a region of interest and eventual overlays more straightforward. Here, we present a workflow for 3D CLEM on mouse liver, combining high-resolution confocal microscopy with SBF-SEM. In this workflow, we have made use of two types of landmarks: (1) near infrared laser branding marks to find back the region imaged in LM in the electron microscope and (2) landmarks present in the tissue but independent of the cell or structure of interest to make overlay images of LM and EM data. Using this approach, we were able to make accurate 3D-CLEM overlays of liver tissue and correlate the fluorescent signal to the ultrastructural detail provided by the electron microscope. This workflow can be adapted for other dense cellular tissues and thus act as a guide for other three-dimensional correlative studies. LAY DESCRIPTION: As cells and tissues exist in three dimensions, microscopy techniques have been developed to image samples, in 3D, at the highest possible detail. In light microscopy, fluorescent probes are used to identify specific proteins or structures either in live samples, (providing dynamic information), or in fixed slices of tissue. A disadvantage of fluorescence microscopy is that only the labeled proteins/structures are visible, while their cellular context remains hidden. Electron microscopy is able to image biological samples at high resolution and has the advantage that all structures in the tissue are visible at nanometer (10 m) resolution. Disadvantages of this technique are that it is more difficult to label a single structure and that the samples must be imaged under high vacuum, so biological samples need to be fixed and embedded in a plastic resin to stay as close to their natural state as possible inside the microscope. Correlative Light and Electron Microscopy aims to combine the advantages of both light and electron microscopy on the same sample. This results in datasets where fluorescent labels can be combined with the high-resolution contextual information provided by the electron microscope. In this study we present a workflow to guide a tissue sample from the light microscope to the electron microscope and image the ultra-structure of a specific cell type in the liver. In particular we focus on the incorporation of fiducial markers during the sample preparation to help navigate through the tissue in 3D in both microscopes. One sample is followed throughout the workflow to visualize the important steps in the process, showing the final result; a dataset combining fluorescent labels with ultra-structural detail.
Topics: Animals; Electrons; Imaging, Three-Dimensional; Liver; Mice; Microscopy, Electron, Scanning; Workflow
PubMed: 33034376
DOI: 10.1111/jmi.12967 -
Nanoscale Advances Feb 2021Direct visualization and imaging of nanomaterials under ambient conditions is of great significance for their characterization and application. In most cases, the... (Review)
Review
Direct visualization and imaging of nanomaterials under ambient conditions is of great significance for their characterization and application. In most cases, the observation of individual nanomaterials usually requires high-resolution electron microscopes under high vacuum. In comparison, an optical microscope is much more convenient due to its facile operation and open space. However, the resolution of optical microscopes is much lower than that of electron microscope-based tools. Therefore, effective visualization and imaging strategies for nanomaterials are required to realize their direct observation, accurate location and controllable manipulation. In this review, we summarized the progress of optical visualization and imaging strategies for nanomaterials in recent years, including vapor-condensation-assisted optical visualization, nanoparticle-assisted optical visualization, substrate-assisted optical visualization and fluorescence visualization, and the applications of these techniques were also introduced. We believe that this review will inspire further improvement in optical visualization of nanomaterials and drive the application of nanomaterials in a broader domain.
PubMed: 36133288
DOI: 10.1039/d0na00945h -
Indian Journal of Ophthalmology Oct 2020Microscopes play an important role in the diagnosis of microorganisms and pathological lesions in ophthalmology guiding us to the appropriate management. The current...
Microscopes play an important role in the diagnosis of microorganisms and pathological lesions in ophthalmology guiding us to the appropriate management. The current trend of collecting samples and examination is mostly laboratory-based which consume time, labor, and are costly. Smartphones are being used in different fields of ophthalmology with great ubiquity. The good quality photographs obtained by smartphones along with the ease of mobility has made it possible to warrant its use in the microscopic world. This article describes a simple novel technique of preparing an intraocular lens system which can be used in conjunction with a smartphone to detect microorganisms and pathological lesions.
Topics: Eye, Artificial; Humans; Lenses, Intraocular; Microscopy; Photography; Smartphone
PubMed: 32971646
DOI: 10.4103/ijo.IJO_2032_19 -
Bio-protocol Oct 2020Cryo-Electron Tomography (cryo-ET) is a method that enables resolving the structure of macromolecular complexes directly in the cellular environment. However, sample...
Cryo-Electron Tomography (cryo-ET) is a method that enables resolving the structure of macromolecular complexes directly in the cellular environment. However, sample preparation for Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes of interest. The PIE-scope can be retrofitted on existing microscopes, although the drawings we provide are meant to work on ThermoFisher DualBeams with small mechanical modifications those can be adapted on other brands.
PubMed: 33659426
DOI: 10.21769/BioProtoc.3768 -
World Neurosurgery Jan 2022Operating microscopes and adjunctive technologies are continually refined to advance microneurosurgical care. How frequently these advances are used is unknown. In the...
BACKGROUND
Operating microscopes and adjunctive technologies are continually refined to advance microneurosurgical care. How frequently these advances are used is unknown. In the present study, we assessed the international adoption of microneurosurgical technologies and discussed their value.
METHODS
A 27-question electronic survey was distributed to cerebrovascular neurosurgeon members of U.S., European, and North American neurosurgical societies and social media networks of cerebrovascular and skull base neurosurgeons. The survey encompassed the surgeons' training background, surgical preferences, and standard microneurosurgical practices.
RESULTS
Of the respondents, 56% (53 of 95) were attendings, 74% (70 of 95) were in their first 10 years of practice, and 67% (63 of 94) practiced at an academic teaching hospital. Vascular, endovascular, and skull base fellowships had been completed by 38% (36 of 95), 27% (26 of 95), and 32% (30 of 95) of the respondents, respectively. Most respondents did not use an exoscope (78%; 73 of 94), a mouthpiece (61%; 58 of 95), or foot pedals (56%; 55 of 94). All 95 respondents used a microscope, and 71 (75%) used Zeiss microscopes. Overall, 57 neurosurgeons (60%) used indocyanine green for aneurysms (n = 54), arteriovenous malformations (n = 43), and dural arteriovenous fistulas (n = 42). Most (80%; 75 of 94) did not use fluorescence. The respondents with a vascular-focused practice more commonly used indocyanine green, Yellow 560 fluorescence, and intraoperative 2-dimensional digital subtraction angiography. The respondents with a skull base-focused practice more commonly used foot pedals and an endoscope-assist device.
CONCLUSIONS
The results from the present survey have characterized the current adoption of operative microscopes and adjunctive technologies in microneurosurgery. Despite numerous innovations to improve the symbiosis between neurosurgeon and microscope, their adoption has been underwhelming. Future advances are essential to improve surgical outcomes.
Topics: Adult; Biomedical Technology; Female; Humans; Internationality; Male; Microsurgery; Middle Aged; Neurosurgeons; Neurosurgical Procedures; Surveys and Questionnaires
PubMed: 34687936
DOI: 10.1016/j.wneu.2021.10.128