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Confocal Microscope

Confocal microscopy is a powerful imaging technique that uses a focused laser beam to scan samples point-by-point, generating high-resolution, 3D images.
This approach enhances contrast and resolution compared to traditional widefield microscopy, making it a valuable tool for biological and materials research.
Confocal microscopes utilize a pinhole aperture to reject out-of-focus light, allowing for improved optical sectioning and the visualization of fine structural details.
With its ability to capture detailed, real-time images of living specimens, confocal microscopy has become indispensible for a wide range of applications, including cell biology, neuroscience, and tissue enginering.
Researchers can leverag the power of confocal imaging to gain new insights into complex biological processes and structures at the subcellular level.

Most cited protocols related to «Confocal Microscope»

1
Single-Molecule RNA FISH of Mouse Embryos
Images were kindly provided by Javier Frias Aldeguer and Nicolas Rivron of Hubrecht Institute for Developmental Biology and Stem Cell Research and Li Linfeng of MERLN Institute for Technology-Inspired Regenerative Medicine. As per Rivron and colleagues [33 (link)], mouse embryos (3.5 dpc) were fixed right after isolation from the mother’s uterus. Fixation was performed using 4% PFA in RNAse-free PBS containing 1% acetic acid. ViewRNA ISH Cell Assay kit (cat# QVC0001) was used for performing smFISH on the embryos. The protocol includes steps of permeabilization and protease treatment as well as probes, preamplifier, amplifier, and label hybridizations. Embryos were then mounted in Slowfade reagent (Thermofisher cat# S36937) and directly imaged in a PerkinElmer Ultraview VoX spinning disk microscope in confocal mode by using a 63×/1.40 NA oil immersion lens.
McQuin C., Goodman A., Chernyshev V., Kamentsky L., Cimini B.A., Karhohs K.W., Doan M., Ding L., Rafelski S.M., Thirstrup D., Wiegraebe W., Singh S., Becker T., Caicedo J.C, & Carpenter A.E. (2018). CellProfiler 3.0: Next-generation image processing for biology. PLoS Biology, 16(7), e2005970.
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Publication 2018
2-5A-dependent ribonuclease Acetic Acids Biological Assay Cells Crossbreeding Embryo isolation Lens, Crystalline Microscopy Mothers Mus Peptide Hydrolases Submersion Uterus
2
Intracellular Labeling of Cortical Neurons
Four 9 month old male mice (C57Bl/SJL) were used. Animals were anesthetized with choral hydrate (15% aqueous solution, i.p.) and were perfused transcardially with 4% paraformaldehyde and 0.125% glutaraldehyde in phosphate buffer saline (PBS; pH 7.4). The brains were then carefully removed from the skull and postfixed for 6 hours. All procedures were conducted in accordance with the National Institute of Health Guide for the Care and Use of Laboratory Animals and were approved by the Mount Sinai School of Medicine Institutional Animal Care and Uses Committee.
For intracellular injections, brains were coronally sectioned at 200 µm on a Vibratome (Leica, Nussloch, Germany). The sections were then incubated in 4,6-diamidino-2-phenylindole (DAPI; Sigma, St. Louis, MO, USA), a fluorescent nucleic acid stain, for 5 minutes, mounted on nitrocellulose filter paper and immersed in PBS. Using DAPI as a staining guide, individual layer II/III pyramidal neurons of the frontal cortex were loaded with 5% Lucifer Yellow (Molecular Probes, Eugene, OR, USA) in distilled water under a DC current of 3–8 nA for 10 minutes, or until the dye had filled distal processes and no further loading was observed [45] (link), [49] (link). Tissue slices were then mounted and coverslipped in Permafluor. Dendritic segment and spine imaging was performed using a Zeiss 410 confocal laser scanning microscope (Zeiss, Thornwood, NY, USA) using a 488 nm excitation wavelength, using a 1.4 N.A. Plan-Apochromat 100× objective with a working distance of 170 µm and a 5× digital zoom. After gain and offset settings were optimized, segments were digitally imaged at 0.1 µm increments, along the optical axis. The confocal stacks were then deconvolved with AutoDeblur (MediaCybernetics, Bethesda, MD, USA).
Supporting Information is available online (Box S1)
Rodriguez A., Ehlenberger D.B., Dickstein D.L., Hof P.R, & Wearne S.L. (2008). Automated Three-Dimensional Detection and Shape Classification of Dendritic Spines from Fluorescence Microscopy Images. PLoS ONE, 3(4), e1997.
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Publication 2008
Animals Animals, Laboratory Brain Buffers Cranium DAPI Dendrites Epistropheus Fingers Glutaral Lobe, Frontal lucifer yellow Males Mice, House Microscopy, Confocal Molecular Probes Nitrocellulose Nucleic Acids paraform Phosphates Protoplasm Pyramidal Cells Saline Solution Stains Tissues Vertebral Column Vision
3
Quantifying Cell Cycle Protein Levels
Cells were grown on Histogrip (Invitrogen) coated glass coverslips and fixed using ice-cold 100% methanol (β-tubulin) or with 3.7% formaldehyde diluted in PBS with 0.5% Triton X-100 for 10 min (Mad2, pSerCdk, Lamin A/C, Plk1, cyclin B1, and securin). All cells were washed and then blocked (3% BSA, 0,1% Tween 20 in PBS) for 30 min. Cells were incubated with primary antibodies were incubated for 2 h at room temperature in blocking solution. DNA was stained with DAPI. For Lamin A/C staining a Leica DM6000 SP8 confocal with a 63× lens was used. All other images were captured using Leica DM5500 microscope coupled with a Coolsnap HQ2 camera, using a Leica 100× or 40× APO 1.4 lens, powered by Leica LAS AF v3 software. To quantify pSer-CDK, cyclin B and secruin levels in cells, a single in-focus plane was acquired. Using ImageJ (v1.48, NIH), an outline was drawn around each cell and circularity, area, mean fluorescence measured, along with several adjacent background readings. The total corrected cellular fluorescence (TCCF) = integrated density – (area of selected cell × mean fluorescence of background readings), was calculated. This TCCF was then equalized against the mean TCCF of neighboring interphase cells in the same field of view, with results presented as fold increase over interphase levels. Box plots and statistical analysis (2-sided unpaired Student t tests) were performed using GraphPad Prism 5. For all other images, 0.3 µm z-sections were taken, de-convolved, and displayed as 2D maximum projections using ImageJ. False coloring and overlays were performed using Adobe Photoshop CS5 software.
McCloy R.A., Rogers S., Caldon C.E., Lorca T., Castro A, & Burgess A. (2014). Partial inhibition of Cdk1 in G2 phase overrides the SAC and decouples mitotic events. Cell Cycle, 13(9), 1400-1412.
Publication 2014
Antibodies Cells Cold Temperature Cyclin B Cyclin B1 DAPI Fluorescence Formaldehyde Interphase Lens, Crystalline LMNA protein, human Methanol Microscopy PLK1 protein, human prisma PTTG1 protein, human Student Triton X-100 Tubulin Tween 20
4
Comprehensive Neuronal Tracing and Characterization
For in situ hybridization analysis, cryostat sections were hybridized using digoxigenin-labeled probes [45 (link)] directed against mouse TrkA or TrkB, or rat TrkC (gift from L. F. Parada). Antibodies used in this study were as follows: rabbit anti-Er81 [14 (link)], rabbit anti-Pea3 [14 (link)], rabbit anti-PV [14 (link)], rabbit anti-eGFP (Molecular Probes, Eugene, Oregon, United States), rabbit anti-Calbindin, rabbit anti-Calretinin (Swant, Bellinzona, Switzerland), rabbit anti-CGRP (Chemicon, Temecula, California, United States), rabbit anti-vGlut1 (Synaptic Systems, Goettingen, Germany), rabbit anti-Brn3a (gift from E. Turner), rabbit anti-TrkA and -p75 (gift from L. F. Reichardt), rabbit anti-Runx3 (Kramer and Arber, unpublished reagent), rabbit anti-Rhodamine (Molecular Probes), mouse anti-neurofilament (American Type Culture Collection, Manassas, Virginia, United States), sheep anti-eGFP (Biogenesis, Poole, United Kingdom), goat anti-LacZ [14 (link)], goat anti-TrkC (gift from L. F. Reichardt), and guinea pig anti-Isl1 [14 (link)]. Terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) to detect apoptotic cells in E13.5 DRG on cryostat sections was performed as described by the manufacturer (Roche, Basel, Switzerland). Quantitative analysis of TUNEL+ DRG cells was performed essentially as described [27 (link)]. BrdU pulse-chase experiments and LacZ wholemount stainings were performed as previously described [46 (link)]. For anterograde tracing experiments to visualize projections of sensory neurons, rhodamine-conjugated dextran (Molecular Probes) was injected into single lumbar (L3) DRG at E13.5 or applied to whole lumbar dorsal roots (L3) at postnatal day (P) 5 using glass capillaries. After injection, animals were incubated for 2–3 h (E13.5) or overnight (P5). Cryostat sections were processed for immunohistochemistry as described [14 (link)] using fluorophore-conjugated secondary antibodies (1:1,000, Molecular Probes). Images were collected on an Olympus (Tokyo, Japan) confocal microscope. Images from in situ hybridization experiments were collected with an RT-SPOT camera (Diagnostic Instruments, Sterling Heights, Michigan, United States), and Corel (Eden Prairie, Minnesota, United States) Photo Paint 10.0 was used for digital processing of images.
Hippenmeyer S., Vrieseling E., Sigrist M., Portmann T., Laengle C., Ladle D.R, & Arber S. (2005). A Developmental Switch in the Response of DRG Neurons to ETS Transcription Factor Signaling. PLoS Biology, 3(5), e159.
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Publication 2005
Anabolism Animals Antibodies Apoptosis Bromodeoxyuridine Calbindins Calretinin Capillaries Cavia Cells Diagnosis Digoxigenin DNA Nucleotidylexotransferase Domestic Sheep Goat Immunohistochemistry In Situ Hybridization In Situ Nick-End Labeling LacZ Genes Lumbar Region Mice, House Microscopy, Confocal Molecular Probes Neurofilaments Neuron, Afferent Pulse Rate Rabbits Rhodamine rhodamine dextran Root, Dorsal Staining transcription factor PEA3 tropomyosin-related kinase-B, human
5
Optical Control and Recording of Neurons
See Supplementary Methods for detailed methods. Constructs with Arch, Mac, and Halo are available at http://syntheticneurobiology.org/protocols. In brief, codon-optimized genes were synthesized by Genscript and fused to GFP in lentiviral and mammalian expression vectors as used previously5 (link),23 (link) for transfection or viral infection of neurons. Primary hippocampal or cortical neurons were cultured and then transfected with plasmids or infected with viruses encoding for genes of interest, as described previously5 (link). Images were taken using a Zeiss LSM 510 confocal microscope. Patch clamp recordings were made using glass microelectrodes and a Multiclamp 700B/Digidata electrophysiology setup, using appropriate pipette and bath solutions for the experimental goal at hand. Neural pH imaging was done using carboxy-SNARF-1-AM ester (Invitrogen). Cell health was assayed using Trypan blue staining (Gibco). HEK cells were cultured and patch clamped using standard protocols. Mutagenesis was performed using the QuikChange kit (Stratagene). Computational modelling of light propagation was done with Monte Carlo simulation with MATLAB. In vivo recordings were made on headfixed awake mice, which were surgically injected with lentivirus, and implanted with a headplate as described before23 (link). Glass pipettes attached to laser-coupled optical fibers were inserted into the brain, to record neural activity during laser illumination in a photoelectrochemical artifact-free way. Data analysis was performed using Clampfit, Excel, Origin, and MATLAB. Histology was performed using transcardial formaldehyde perfusion followed by sectioning and subsequent confocal imaging.
Chow B.Y., Han X., Dobry A.S., Qian X., Chuong A.S., Li M., Henninger M.A., Belfort G.M., Lin Y., Monahan P.E, & Boyden E.S. (2010). High-Performance Genetically Targetable Optical Neural Silencing via Light-Driven Proton Pumps. Nature, 463(7277), 98-102.
Publication 2009
Bath Brain carboxy-seminaphthorhodaminefluoride Cells Cloning Vectors Codon Cortex, Cerebral Esters Formaldehyde Genes Lentivirus Light Mammals Microelectrodes Microscopy, Confocal Mus Mutagenesis Nervousness Neurons Operative Surgical Procedures Perfusion Plasmids Transfection Trypan Blue Virus Virus Diseases

Most recents protocols related to «Confocal Microscope»

1
Detailed Confocal Microscopy Imaging Protocol
All images were captured by a Leica STELLARIS confocal microscopy and analyzed by the LAS X Life Science Microscope Software.
Wei Z., Hao C., Radeen K.R., Srinivasagan R., Chen J.K., Sharma S., McGee-Lawrence M.E., Hamrick M.W., Monnier V.M, & Fan X. (2024). Prevention of age-related truncation of γ-glutamylcysteine ligase catalytic subunit (GCLC) delays cataract formation. Science Advances, 10(17), eadl1088.
Publication 2024
2
Deciphering LATS2, SIAH2, CTGF, YAP, and α-SMA Interplay
Tissue sections were stained with antibodies to LATS2 (Novus, USA), SIAH2 (Novus, USA), CTGF (Santa-Cruz, USA), YAP (CST, USA) and α-SMA (Protein-tech, USA) for 14–16 h at 4℃ and followed by staining with Alexa-488 conjugated anti-Rabbit and Alexa-595 conjugated anti-Mouse respectively. 4’, 6-diamidino-2- phenylindole dihydrochloride (DAPI) was used to color nuclei. Confocal images were recorded by FV3000 confocal microscope (Olympus, Japan).
Cheng C., Yang H., Yang C., Xie J., Wang J., Cheng L., He J., Li H., Yuan H., Guo F., Li M, & Liu S. (2024). LATS2 degradation promoted fibrosis damage and rescued by vitamin K3 in lupus nephritis. Arthritis Research & Therapy, 26, 64.
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Publication 2024
3
Minocycline Ocular Uptake Dynamics
The intrinsic fluorescence intensity of minocycline was observed using a fluorescence microscope, with cell nuclei being labeled with DAPI. MINO@PLGA and minocycline were smeared on slides and calibrated under a fluorescence microscope. Rats were then grouped by various time points and administered the drug once. At the designated time points, rat eyeballs and surrounding tissues were harvested and processed into frozen sections. The frozen sections were fixed in cold acetone for 10 min, then washed, and subsequently mounted using a DAPI medium. Fluorescent images of the cornea were captured using a confocal microscope, employing the specified parameters.
Li Z., Huang W., Zhang M., Huo Y., Li F., Song L., Wu S., Yang Q., Li X., Zhang J., Yang L., Hao J, & Kang L. (2024). Minocycline-loaded nHAP/PLGA microspheres for prevention of injury-related corneal angiogenesis. Journal of Nanobiotechnology, 22, 134.
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Publication 2024
4
Acridine Orange Staining for Confocal Imaging
Not available on PMC !
Cells were cultured on glass-based Petri dishes and stained with 10 µm acridine orange, the signal was collected by exciting samples with 405 nm and 488 nm lasers, and emission was collected at 542 nm (represented by red) and 494 nm (represented by green), respectively and imaged with confocal scanning microscope (CLSM, LSM 710, Carl Zeiss, Germany)
Bashir S., Moghazy G.M.E., Aal M.H.A., & El-Jakee J. (2024). Prebiotic and Probiotic: A Promising Tool in Human Colon Cancer Prevention. Egyptian Journal of Veterinary Science, 55(3), 817-824.
Publication 2024
5
Mitochondrial Depolarization and ROS Imaging
ΔφM and reactive oxygen species (ROS) production were observed using confocal microscopy (Nikon Instruments A1 Confocal Laser Microscope Series With NIS‐Elements C Software). This assessment was conducted considering mitochondrial depolarization as the initial mitochondrial response to drug‐induced stress.
27 (link) The cells were placed in glass‐bottom dishes with and without 100 nM doxorubicin and treated for 6 h. They were then stained with tetramethyl rhodamine, methyl ester (TMRM) (400 nM, Marker Gene Technologies INC, USA), and Mito Marker Green (MTG) (120 nM, Marker Gene Technologies, Inc, USA) for 30 min. Additionally, The CellROX ROS Detection Kit (1:1000, ab186029, Abcam) was incubated for 45 min in a cell culture incubator. Hoechst 33342 (1:10,000, H3570, Thermo Scientific, USA) was added and incubated for 10 min. The excitation wavelengths for TMRM, MTG, ROS, and Hoechst 33342 were 548, 490, 650, and 361 nm, respectively, while their emission wavelengths were 574, 516, 675, and 486 nm, respectively.
Wang Y., Harada‐Shoji N., Kitamura N., Yamazaki Y., Ebata A., Amari M., Watanabe M., Miyashita M., Tada H., Abe T., Suzuki T., Gonda K, & Ishida T. (2024). Mitochondrial dynamics as a novel treatment strategy for triple‐negative breast cancer. Cancer Medicine, 13(2), e6987.
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Publication 2024

Top products related to «Confocal Microscope»

1
Lsm 710 by Zeiss
Sourced in Germany, United States, United Kingdom, Japan, Switzerland, France, China, Canada, Italy, Spain, Singapore, Austria, Hungary, Australia
The LSM 710 is a laser scanning microscope developed by Zeiss. It is designed for high-resolution imaging and analysis of biological and materials samples. The LSM 710 utilizes a laser excitation source and a scanning system to capture detailed images of specimens at the microscopic level. The specific capabilities and technical details of the LSM 710 are not provided in this response to maintain an unbiased and factual approach.
2
4 6 diamidino 2 phenylindole (dapi) by Merck Group
Sourced in United States, Germany, Japan, United Kingdom, China, Italy, Sao Tome and Principe, France, Macao, Canada, Switzerland, Spain, Australia, Denmark, India, Poland, Israel, Belgium, Sweden, Ireland, Netherlands, Panama, Brazil, Portugal, Czechia, Puerto Rico, Austria, Hong Kong, Singapore
DAPI is a fluorescent dye that binds strongly to adenine-thymine (A-T) rich regions in DNA. It is commonly used as a nuclear counterstain in fluorescence microscopy to visualize and locate cell nuclei.
3
4 6 diamidino 2 phenylindole (dapi) by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, Japan, China, Canada, Italy, Australia, France, Switzerland, Spain, Belgium, Denmark, Panama, Poland, Singapore, Austria, Morocco, Netherlands, Sweden, Argentina, India, Finland, Pakistan, Cameroon, New Zealand
DAPI is a fluorescent dye used in microscopy and flow cytometry to stain cell nuclei. It binds strongly to the minor groove of double-stranded DNA, emitting blue fluorescence when excited by ultraviolet light.
4
Lsm 710 confocal microscope by Zeiss
Sourced in Germany, United States, United Kingdom, France, Switzerland, Canada, Japan, Singapore, Italy
The ZEISS LSM 710 is a confocal laser scanning microscope. It enables high-resolution imaging of samples by using a focused laser beam to scan the specimen point-by-point, and then detecting the emitted fluorescence or reflected light. The microscope is designed to provide researchers with a versatile and reliable tool for a wide range of imaging applications.
5
Lsm 700 by Zeiss
Sourced in Germany, United States, Japan, Canada, United Kingdom, Switzerland, France, Italy, China, Denmark, Australia, Austria, Slovakia, Morocco
The LSM 700 is a versatile laser scanning microscope designed for high-resolution imaging of samples. It provides precise control over the illumination and detection of fluorescent signals, enabling detailed analysis of biological specimens.
6
Lsm 880 by Zeiss
Sourced in Germany, United States, United Kingdom, Japan, China, Switzerland, France, Austria, Canada, Australia
The LSM 880 is a laser scanning confocal microscope designed by Zeiss. It is a versatile instrument that provides high-resolution imaging capabilities for a wide range of applications in life science research.
7
Fv1000 by Olympus
Sourced in Japan, United States, Germany, China, France, United Kingdom, Canada
The FV1000 is a confocal laser scanning microscope designed for high-resolution imaging of biological samples. It features a modular design and advanced optics to provide clear, detailed images of cellular structures and processes.
8
Lsm 780 by Zeiss
Sourced in Germany, United States, Japan, France, China, Canada, United Kingdom, Switzerland, Singapore, Italy, Panama, India
The LSM 780 is a laser scanning microscope developed by Zeiss. It is designed for high-resolution imaging and analysis of biological samples. The instrument utilizes advanced confocal technology to provide detailed, three-dimensional images of specimens.
9
Lsm 700 confocal microscope by Zeiss
Sourced in Germany, United States, France, Canada, Japan, United Kingdom, Sweden, Italy, Hungary, Switzerland, Denmark, Albania
The LSM 700 is a confocal microscope manufactured by Zeiss. It is designed to provide high-resolution imaging of samples by utilizing laser scanning technology. The core function of the LSM 700 is to produce detailed, three-dimensional images of various specimens through optical sectioning.
10
Sp8 confocal microscope by Leica
Sourced in Germany, United States, Japan, Italy, France, United Kingdom, China, Switzerland, Canada, Portugal
The Leica SP8 confocal microscope is a high-performance imaging system designed for advanced microscopy applications. It features a state-of-the-art confocal architecture that enables high-resolution, real-time imaging of fluorescently labeled samples. The SP8 offers precise control over laser excitation, detector settings, and optical parameters to optimize image quality and data acquisition.

More about "Confocal Microscope"

Confocal Microscopy: Illuminating the Subcellular World Confocal microscopy, a revolutionary imaging technique, has become an indispensable tool in the world of biological and materials research.
This powerful method, also known as laser scanning confocal microscopy (LSCM) or laser confocal scanning microscopy (LCSM), utilizes a focused laser beam to scan samples point-by-point, generating high-resolution, three-dimensional (3D) images with unparalleled clarity and contrast.
Unlike traditional widefield microscopy, confocal microscopy enhances optical sectioning and rejects out-of-focus light, allowing researchers to visualize fine structural details with unprecedented precision.
This approach has transformed our understanding of complex biological processes and structures at the subcellular level, making it a vital instrument in fields such as cell biology, neuroscience, and tissue engineering.
The key to confocal microscopy's success lies in its use of a pinhole aperture, which filters out-of-focus light, resulting in sharper, more detailed images.
This technique, coupled with advanced software and hardware, enables real-time imaging of living specimens, opening up new frontiers in the study of dynamic cellular events.
Researchers have a wide range of confocal microscope models to choose from, including the popular LSM 710, LSM 700, LSM 880, FV1000, and SP8 systems.
These state-of-the-art instruments, equipped with versatile features such as multi-channel detection, spectral imaging, and DAPI (4',6-diamidino-2-phenylindole) staining, provide unparalleled flexibility and precision in their applications.
By leveraging the power of confocal imaging, scientists can gain deeper insights into complex biological structures and processes, from the intricate dance of cellular organelles to the delicate networks of neural connections.
This technology has become an indispensable tool in the quest to unlock the secrets of the living world, driving forward the boundaries of scientific discovery.