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> Procedures > Laboratory Procedure > Light Microscopy

Light Microscopy

Light microscopy is a powerful technique used to visualize and analyze microscopic biological structures and processes.
This versatile methodology employs visible light to magnify samples, enabling researchers to study cellular, subcellular, and tissue-level details.
Light microscopy encompasses a wide range of modalities, including brightfield, phase contrast, fluorescence, and differential interference contrast, each offering unique capabilities for different applications.
Through the integration of advanced imaging systems and computational analysis, light microscopy continues to evolve, providing scientists with unprecedented insights into the complex world of the microscopic realm.
Whether investigating cellular dynamics, tissue architecture, or molecular interactions, light microscopy remains an indispensable tool in the field of biomedical research.

Most cited protocols related to «Light Microscopy»

1
Immunohistochemical Staining of Tumor Tissue
Standard IHC protocol was followed to stain the tumor tissue samples using the mouse monoclonal antibody against hNIS (human Sodium Iodide Symporter) (Abcam, ab17795), ER (Estrogen Receptor) (Abcam, ab16660, ab288). Briefly, 5 µm sized paraffin embedded tissue sections were de-paraffinized with xylene and endogenous peroxidase activity was quenched with 3% H2O2 in methanol for 30 minutes in the dark. Tissue sections were dehydrated through graded alcohols and subjected to antigen retrieval using 10mM sodium citrate. Sections were washed with TBST (Tris Borate Saline Tween-20) and then blocked with 5% BSA (Bovine Serum Albumin) for one hour. Slides were incubated with the respective mouse monoclonal primary antibody diluted with TBS. Slides were then washed for 5 minutes in TBST and incubated for 1 hour with the respective HRP (Horse Raddish Peroxidase) conjugated anti-mouse secondary antibody diluted with TBS in a ratio of 1∶200. After washing, slides were incubated with DAB (3,3′-diaminobenzidine tetrahydrochloride) (Sigma) and immediately washed under tap water after color development. Slides were then counter stained with hematoxylin. Slides were mounted with DPX (dibutyl phthalate xylene) and were then observed under a light microscope (Carl Zeiss).
Varghese F., Bukhari A.B., Malhotra R, & De A. (2014). IHC Profiler: An Open Source Plugin for the Quantitative Evaluation and Automated Scoring of Immunohistochemistry Images of Human Tissue Samples. PLoS ONE, 9(5), e96801.
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Publication 2014
Antibodies, Anti-Idiotypic Antigens Borates Equus caballus estrogen receptor alpha, human Ethanol Homo sapiens Light Microscopy Methanol Monoclonal Antibodies Mus Neoplasms Paraffin Peroxidase Peroxide, Hydrogen Phthalate, Dibutyl Saline Solution Serum Albumin, Bovine SLC5A5 protein, human Sodium Citrate Stains Tissues Tromethamine Tween 20 Xylene
2
Efficient Arabidopsis Protoplast Isolation
Leaves (width: 2 cm, length: 5 cm in optimal light condition; width: 0.5 cm; length: 2.5 cm in low light conditions) were collected from 3 to 5-week-old plants grown under optimal light (ca. 150 μE·m-2·s-1) or low light (ca. 50·μE m-2·s-1) conditions. Arabidopsis protoplasts were isolated in two ways. First, to recreate the current technique, protoplasts were made according to the procedure of Yoo et al. [4 (link)]. Second, in a new technique, selected leaves were used in a 'Tape-Arabidopsis Sandwich' experiment. The upper epidermal surface was stabilized by affixing a strip of Time tape (Time Med, Burr Ridge, IL) while the lower epidermal surface was affixed to a strip of Magic tape (3 M, St. Paul, MN). The Magic tape was then carefully pulled away from the Time tape, peeling away the lower epidermal surface cell layer. The peeled leaves (7 to 10 optimal-light-growth leaves, about 1-2 g, up to 5 g), still adhering to the Time tape, were transferred to a Petri dish containing 10 mL of enzyme solution [1% cellulase 'Onozuka' R10 (Yakult, Tokyo, Japan), 0.25% macerozyme 'Onozuka' R10 (Yakult), 0.4 M mannitol, 10 mM CaCl2, 20 mM KCl, 0.1% BSA and 20 mM MES, pH 5.7]. The leaves were gently shaken (40 rpm on a platform shaker) in light for 20 to 60 min until the protoplasts were released into the solution. The protoplasts were centrifuged at 100 × g for 3 min in an Eppendorff A-4-44 rotor (Hamburg, Germany), washed twice with 25 mL of pre-chilled modified W5 solution (154 mM NaCl, 125 mM CaCl2, 5 mM KCl, 5 mM glucose, and 2 mM MES, pH 5.7) and incubated on ice for 30 min. During the incubation period, protoplasts were counted using a hemocytometer under a light microscope. The protoplasts were then centrifuged and resuspended in modified MMg solution (0.4 M mannitol, 15 mM MgCl2, and 4 mM MES, pH 5.7) to a final concentration of 2 to 5 × 105 cells/mL.
Wu F.H., Shen S.C., Lee L.Y., Lee S.H., Chan M.T, & Lin C.S. (2009). Tape-Arabidopsis Sandwich - a simpler Arabidopsis protoplast isolation method. Plant Methods, 5, 16.
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Publication 2009
Arabidopsis Cells Cellulase Enzymes Epidermal Cells Epidermis Glucose Hyperostosis, Diffuse Idiopathic Skeletal Light Light Microscopy Magnesium Chloride Mannitol Plants Protoplasts Sodium Chloride
3
Immunophenotypic Analysis of Cell Viability
After cells were counted, and 2x106 cells per sample were stained with Aqua Live/Dead viability dye (Life Technologies) according the manufacturer’s instructions. Cells were then incubated in blocking solution containing 5% normal mouse serum, 5% normal rat serum, and 1% FcBlock (eBiosciences, San Diego, CA) in PBS and then stained with a standard panel of immunophenotyping antibodies (See S2 Table for a list of antibodies, clones, fluorochromes, manufacturers, and concentrations) for 30 minutes at room temperature. After staining, cells were washed and fixed with 0.4% paraformaldehyde in PBS. Data was acquired with a BD LSRII flow cytometer using BD FACSDiva software (BD Bioscience). Compensation was performed on the BD LSRII flow cytometer at the beginning of each experiment. Data were analyzed using Flowjo v10. Cell sorting for cytospins was performed on a BD Aria II. The collected cells were stained with a Jenner-Giemsa Stain Kit (ENG Scientific Inc, Clifton, NJ) and examined by light microscopy.
Yu Y.R., O’Koren E.G., Hotten D.F., Kan M.J., Kopin D., Nelson E.R., Que L, & Gunn M.D. (2016). A Protocol for the Comprehensive Flow Cytometric Analysis of Immune Cells in Normal and Inflamed Murine Non-Lymphoid Tissues. PLoS ONE, 11(3), e0150606.
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Publication 2016
Antibodies Cells Clone Cells Fluorescent Dyes Light Microscopy Mus NRG1 protein, human paraform Serum Stain, Giemsa Stains
4
Quantitative Landmark Analysis of Micrasterias
The studied dataset comprised 68 mature cells taken from CAUP K608, a clonal strain of Micrasterias compereana. This strain, which has been used as a holotype for the taxonomic description of the species by Neustupa et al. [46 (link)], was originally isolated in 2011 from oligotrophic peaty pools near Étang Hardy, Aquitaine, France (43°43′08.60″N, 01°22′09.42″W). It was cultivated in 250 ml Erlenmeyer flasks with approximately 125 ml of the MES-buffered DY IV liquid medium at 22 °C and illuminated at 40 μmol photons m−2 s−1 with 18 W cool fluorescent tubes (Philips TLD 18 W/33), at a light:dark (L:D) regime of 12:12 h.
The cells were photographed at 200× magnification on an Olympus BX51 light microscope with Olympus DP27 digital photographic equipment. In total, 208 structurally corresponding landmarks were depicted on the front-view images of the cells (Fig. 1b, Additional file 1) using TpsDig software, ver. 2.15 [47 ]. To assess the measurement error, all landmarks were digitised twice. In the first digitisation, the landmarks were registered clockwise starting from the left margin of the cellular isthmus. Conversely, the second digitisation proceeded counter clockwise from the same starting point and the landmarks were relabelled to match the labels of the first digitisation.
, & Neustupa J. (2017). Asymmetry and integration of cellular morphology in Micrasterias compereana. BMC Evolutionary Biology, 17, 1.
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Publication 2017
Cells Clone Cells Fingers Light Microscopy Micrasterias Mucolipidosis Type IV Strains
5
Comparative Analysis of Super Black Plumage
Five bird species with profoundly black plumage and two species with normal black plumage were identified by visual observation of museum study skins from the Yale Peabody Museum (YPM), Harvard Museum of Comparative Zoology (MCZ), American Museum of Natural History (AMNH), and the University of Kansas Biodiversity Institute (KU). Details of the specimens and plumage patches studied are summarized in Supplementary Table 1. To the human observer, super black plumage had a strongly matte appearance with so little specular reflectance that it was difficult to focus on the surface of the plumage and distinguish individual feathers. The species with normal black plumage lacked any conspicuous glossy specular highlights. Individual contour feathers were sampled from museum skins for scanning electron microscopy (SEM) and synchrotron-radiation X-ray microtomograhy (nano-CT). We could not obtain SEM of Lophorina superba back feathers or CT scans for Lophorina superba back and display cape feathers due to availability of material. Visual inspection of the Lophorina back plumage using a light microscope confirmed that the barbules have normal morphology, without the modified barbule arrays present in super black feathers.
McCoy D.E., Feo T., Harvey T.A, & Prum R.O. (2018). Structural absorption by barbule microstructures of super black bird of paradise feathers. Nature Communications, 9, 1.
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Publication 2018
Aves Feathers Homo sapiens Light Microscopy Radiation Radiography Scanning Electron Microscopy Skin X-Ray Computed Tomography

Most recents protocols related to «Light Microscopy»

1
pHLIP-Mediated Tumor-Targeted Liposome Delivery
Not available on PMC !

Example 4

Liposomes, containing Rho-PE lipids, were given as a single intra-tumoral injection into mice with tumors established by subcutaneous injection of HeLa-GFP cancer cells. Mice were sacrificed at 24 hours post-injection, and tumors were collected. Whole-body and tumor images were taken on Kodak in vivo imaging system. As shown in FIG. 13, pHLIP promoted liposome uptake in low pH extracellular environment of tumors, following IV injection of the fluorescent- and gold-containing liposomes.

HeLa-GFP cells were incubated with pHLIP-nanogold and nanogold particles at neutral and low pHs, washed, fixed and enhanced by silver then visualized under light microscope. The highest uptake was observed at low pH in presence of pHLIP (FIG. 17A). Tumor sections collected from mice received single iv injection of pHLIP-nanogold and nanogold particles were treated with silver enhancement solution and visualized under the microscope. Nanogold particles delivered to tumor by pHLIP were localized on cancer cells identified by GFP fluorescence (FIG. 17B).

These data indicate that pHLIP-liposomes demonstrate enhanced uptake by cells in environments characterized by low pH (pH<7) compared to liposomes that do not contain pHLIP.

US11857509B2. Liposome compositions and methods of use thereof (2024-01-02). University of Rhode Island Board of Trustees [US], Yale University [US]. Inventors: Yana K. Reshetnyak [US], Oleg A. Andreev [US], Donald M. Engelman [US].
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Patent 2024
Cells Fluorescence Gold HeLa Cells Human Body Light Microscopy Lipids Liposomes Malignant Neoplasms Microscopy Mus Neoplasms Silver Subcutaneous Injections
2
Simulated Hyperspectral Test Chart for Microscopy
Not available on PMC !

Example 13

To account for the Poisson noise and detector noise contributed by optical microscopy, we generated a simulated hyperspectral test chart starting from real imaging data with a size of x: 300 pixels, y: 300 pixels, and lambda: 32 channels. S1, S2 and S3 spectra were acquired respectively from zebrafish embryos labeled only with CFP, YFP, and RFP, where the spectrum in FIG. 24a is identical to the center cell of the test chart FIG. 24d. In each cell three spectra are represented after shifting the maxima by d1 or d2 nanometers with respect to S2. Each cell has its corresponding spectra of S1, S2, and S3 (FIG. 33).

US11869176B2. Hyperspectral imaging system (2024-01-09). UNIVERSITY OF SOUTHERN CALIFORNIA [US]. Inventors: Wen Shi [US], Eun Sang Koo [US], Scott E. Fraser [US], Francesco Cutrale [US].
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Patent 2024
Cells Embryo Light Microscopy Zebrafish
3
Candidiasis Microscopic Profiles
Candidiasis profiles are defined as the characteristic appearance of the wet mount of candida samples under a high-power field light microscopy. The candidiasis profiles under consideration were epithelial cells (ECs), pus cells (PCs), red blood cells (RBCs) and Candida albicans positive in a wet mount preparation. The patients’ demographic characteristics including patient age, sex, the type of samples and the year of diagnosis were also obtained.
Asare K.K., Bentil H.A., Gyesi E., Amoah S., Bentsi-Enchill F, & Opoku Y.K. (2023). Candidiasis profile at the outpatient department of the university of cape coast hospital in the central region of Ghana: a retrospective study. BMC Women's Health, 23, 101.
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Publication 2023
Candida Candida albicans Candidiasis Cells Diagnosis Epithelial Cells Erythrocytes Light Microscopy Patients
4
Estrous Cycle Assessment in Female Mice
Estrous cycles were assessed at the same time every day during behavioral study performance day following published protocols [53 (link), 54 (link)]. Female mice were properly handled to minimize stress, by gently lifting the animal by the base of tail a plastic pipette filled with about 1 ml of PBS was placed on the tip of the vagina and flushed 5 times with same PBS to allow proper collection of samples for vaginal cytology. Sample was then smeared on appropriate labeled microscope slides and after 1 h of drying time, a crystal violet (0.1%) staining was performed on slides. After drying, the slides were observed under a light microscope to visualize cells. Images were obtained using NIS Elements imaging software version 4.0.
Archie S.R., Sifat A.E., Zhang Y., Villalba H., Sharma S., Nozohouri S, & Abbruscato T.J. (2023). Maternal e-cigarette use can disrupt postnatal blood-brain barrier (BBB) integrity and deteriorates motor, learning and memory function: influence of sex and age. Fluids and Barriers of the CNS, 20, 17.
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Publication 2023
Animals Cells Cytological Techniques Estrous Cycle Females Light Microscopy Microscopy Mus Specimen Collection Tail Vagina Violet, Gentian
5
Pancreas Clearing and Imaging Protocol
The pancreas was microdissected from control-Ai14 and Isl1CKO-Ai14 mice (postnatal day P0). We used an advanced CUBIC protocol [65 (link)] for tissue clearing to enable efficient imaging by light-sheet microscopy. Briefly, the microdissected tissue was fixed in 4% PFA for 1 h, washed with PBS, and incubated in a clearing solution Cubic 1 for 5 days at 37 ℃. Before immunolabeling, samples were washed in PBT (0.5% Triton-X in PBS) 4 × for 30 min. In addition to tdTomato expression, cleared samples were immunolabeled using different combinations of antibodies (anti-INS, anti-GLP1, and anti-TUBB3). Samples were stored before imaging in Cubic 2 at room temperature. Zeiss Lightsheet Z.1 microscope with illumination objective Lightsheet Z.1 5x/0.1 and detection objective Dry objective Lightsheet Z.1 5x/0.16 was used for imaging at the Light Microscopy Core Facility of the Institute of Molecular Genetics of the Czech Academy of Sciences. IMARIS software v8.1.1 (Bitplane AG, CA, USA) was used for image processing.
Bohuslavova R., Fabriciova V., Lebrón-Mora L., Malfatti J., Smolik O., Valihrach L., Benesova S., Zucha D., Berkova Z., Saudek F., Evans S.M, & Pavlinkova G. (2023). ISL1 controls pancreatic alpha cell fate and beta cell maturation. Cell & Bioscience, 13, 53.
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Publication 2023
Antibodies Cuboid Bone EHMT1 protein, human Light Microscopy Mus Pancreas tdTomato Tissues TUBB3 protein, human

Top products related to «Light Microscopy»

1
Matrigel by BD
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2
Optical microscope by Olympus
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The Olympus optical microscope is a precision instrument designed for detailed observation and analysis of small-scale specimens. It utilizes advanced optical technology to magnify and illuminate samples, enabling users to examine fine details and structures not visible to the naked eye. The core function of this product is to provide a reliable and high-quality platform for microscopic examination across a variety of scientific and research applications.
3
Transwell chamber by Corning
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Transwell chambers are a type of lab equipment used for cell culture and biological assays. They consist of a permeable membrane insert placed inside a well, allowing for the study of cell-cell interactions and the movement of molecules across a barrier. The core function of Transwell chambers is to provide a controlled environment for culturing cells and monitoring their behavior and permeability.
4
Image pro plus 6 by Media Cybernetics
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Image-Pro Plus 6.0 is a comprehensive image analysis software package designed for scientific and industrial applications. It provides a wide range of tools for image capture, enhancement, measurement, analysis, and reporting.
5
Eclipse 80i by Nikon
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The Eclipse 80i is a microscope designed for laboratory use. It features an infinity-corrected optical system and offers a range of illumination options. The Eclipse 80i is capable of various imaging techniques, including phase contrast and brightfield microscopy.
6
Crystal violet by Merck Group
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Crystal violet is a synthetic dye commonly used in laboratory settings. It is a dark purple crystalline solid that is soluble in water and alcohol. Crystal violet has a variety of applications in the field of microbiology and histology, including as a staining agent for microscopy and in the gram staining technique.
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Matrigel by Corning
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Matrigel is a complex mixture of extracellular matrix proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells. It is widely used as a basement membrane matrix to support the growth, differentiation, and morphogenesis of various cell types in cell culture applications.
8
Light microscopy by Olympus
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Light microscopy is an optical imaging technique that uses visible light and a system of lenses to magnify and examine small objects. It allows for the observation and analysis of microscopic structures and details that are not visible to the naked eye.
9
Bx51 light microscope by Olympus
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The Olympus BX51 is a light microscope designed for high-quality observation and imaging. It features a robust optical system, including a Plan-Achromat objective lens, to provide clear and accurate imaging. The BX51 is capable of bright-field, dark-field, and polarized light microscopy techniques.
10
Inverted light microscope by Olympus
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The Inverted Light Microscope is a type of microscope designed for observing specimens from below. It features a reversed orientation, with the light source and condenser located above the stage, and the objective lens and eyepiece below. This configuration allows for the examination of live cell cultures, tissue samples, and other specimens that require a specific orientation or environment.

More about "Light Microscopy"

Discover the power of light microscopy in biomedical research.
This versatile technique employs visible light to magnify and analyze microscopic biological structures and processes, offering researchers unparalleled insights.
From cellular dynamics to tissue architecture, light microscopy encompasses a wide range of modalities, including brightfield, phase contrast, fluorescence, and differential interference contrast, each with unique capabilities for diverse applications.
Complement your light microscopy workflow with cutting-edge tools like Matrigel, Optical microscopes, Transwell chambers, Image-Pro Plus 6.0, Eclipse 80i, and BX51 light microscopes, which can further enhance your experimental capabilities.
Whether you're investigating subcellular interactions, molecular events, or tissue-level details, light microscopy remains an indispensable and evolving tool, constantly pushing the boundaries of scientific discovery.
Streamline your research and unlock new possibilities with the help of AI-driven platforms like PubCompare.ai, which can effortlessly locate the best protocols from literature, preprints, and patents, empowering you to identify the optimal techniques and products for your experiments.
Embrace the power of light microscopy and let it guide you through the complex and fascinating world of the microscopic realm.