Dapi containing mounting media

Manufactured by Vector Laboratories

Sourced in United States, Canada

DAPI-containing mounting media is a solution designed for mounting and preserving fluorescently labeled biological samples on microscope slides. The primary function of this product is to provide a suitable medium for mounting specimens and preserving fluorescent signals, including those from DAPI (4',6-diamidino-2-phenylindole) staining.

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Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (6 mentions) Upright confocal microscopy, by Leica (4 mentions) Image pro software, by Media Cybernetics (3 mentions) Fluorescent microscope, by Leica (3 mentions) Triton x 100, by Merck Group (3 mentions) Lsm 710 confocal laser scanning microscope, by Zeiss (2 mentions) Glucagon, by Agilent Technologies (2 mentions) Tunel technique, by Roche (2 mentions) Fatal plus, by Vortech Pharmaceuticals (2 mentions) Eclipse 80i, by Nikon (2 mentions) Caseviewer software, by 3DHISTECH (2 mentions) Pannoramic 250 scanner, by 3DHISTECH (2 mentions) Apotome microscope, by Zeiss (2 mentions) Fluorescein isothiocyanate (fitc), by Jackson ImmunoResearch (2 mentions) Upright fluorescence microscope, by Leica (2 mentions) Ab7952, by Abcam (2 mentions) Bx ucb fluorescent microscope, by Olympus (2 mentions) Nl006, by R&D Systems (2 mentions) Fluorescein conjugated secondary antibody, by Merck Group (2 mentions) Lsm 780, by Zeiss (2 mentions)

Close spelling variants of this product

Vectashield antifade mounting media containing 4′,6-diamidino-2-phenylindole (DAPI) (2 citations) Vectashield mounting media containing DAPI (17 citations) DAPI-containing fluorescent mounting media (3 citations) DAPI-containing media (12 citations) DAPI mounting media (31 citations) Mounting media (59 citations)

46 protocols using dapi containing mounting media

Imaging of Neuronal and Astrocytic Markers

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Fixed cells
were stained for β₃-tubulin (1:500) and GFAP (1:1000)
using secondary antibodies tagged with fluorescent markers (Alexa
Fluor 488 and 555, 1:300); all antibodies and markers were obtained
from Abcam. Samples were mounted in DAPI-containing mounting media
(Vector Labs, Peterborough). Samples were imaged on a Nikon epifluorescence
microscope fitted with an auto-x-y scanning stage, allowing large-area
stitched image acquisition. All images were taken at ×20 magnification.
Confocal imaging was carried out on a Zeiss LSM 710 confocal laser
scanning microscope fitted with 20× objective. Layer slices were
captured at a resolution of 45 μm with successive layers being
pictured.

Joseph G., Orme R.P., Kyriacou T., Fricker R.A, & Roach P. (2021). Effects of Surface Chemistry Interaction on Primary Neural Stem Cell Neurosphere Responses. ACS Omega, 6(30), 19901-19910.

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Quantifying Pancreatic Islet Composition

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Formalin-fixed pancreatic tissues were embedded in paraffin, sectioned, deparaffinized and rehydrated using standard techniques. For immunofluorescence, sections were incubated overnight at 4 °C with antibodies against insulin (Abcam, Cambridge, UK), glucagon (Dako, Glostrup, Denmark), and/or Ki67 (Abcam, Cambridge, UK), followed by secondary antibodies conjugated to FITC or Cy3 (Life Technologies). DAPI-containing mounting media (Vector Laboratories, Burlingame, CA, USA) was added to coverslips. Apoptotic cells were identified by the TUNEL technique (Roche). Immunofluorescence staining was quantified using ImageJ. For quantification, all the islets embedded in 2 pancreatic sections separated by 200 μm were analyzed, resulting in the counting of at least 500 beta-cells/mouse. For beta-cell area measurements, the percentage of insulin-positive surface area was determined in 8 evenly spaced slices of pancreas using ImageScope (Aperio).

Huerta Guevara A.P., McGowan S.J., Kazantzis M., Stallons T.R., Sano T., Mulder N.L., Jurdzinski A., van Dijk T.H., Eggen B.J., Jonker J.W., Niedernhofer L.J, & Kruit J.K. (2021). Increased insulin sensitivity and diminished pancreatic beta-cell function in DNA repair deficient Ercc1d/− mice. Metabolism: clinical and experimental, 117, 154711.

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Perfusion and Sectioning for GCaMP Imaging

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Mice were deeply anesthetized with FatalPlus^TM (Vortech Pharmaceuticals, Dearborn, MI, USA) and transcardially perfused with 0.01 M PBS. The brains were removed and fixed in 4% paraformaldehyde for 24 h and transferred to 30% sucrose in 0.2 M PBS. After brains were saturated, they were snap-frozen on dry ice and coronal sections, 50 μm thick, were made with a sliding microtome. Sections were stored in 0.2 M PBS, 30% sucrose, and 30% ethylene glycol at −20°C. For view ing, cut sections were washed in PBS, mounted, and intrinsic GCaMP6 fluorescence was examined with epifluorescence microscopy (Nikon Eclipse 80i, Nikon Instruments Inc., Melville, NY, USA). For viewing using confocal microscopy, additional cut sections were washed in PBS, mounted, and coverslipped in DAPI containing mounting media (Vector Laboratories, Burlingame, CA, USA). Fluorescent images were acquired with a Nikon A1-Rsi inverted confocal microscope using a 10x objective. DAPI labeled cells and electrode marker DiI were excited with 405nm and 560nm laser lines, respectively (Nikon A1-Rsi, Nikon Instruments Inc., Melville, NY, USA).

Mitra A., Kraft A., Wright P., Acland B., Snyder A.Z., Rosenthal Z., Czerniewski L., Bauer A., Snyder L., Culver J., Lee J.M, & Raichle M.E. (2018). Spontaneous infra-slow brain activity has unique spatio-temporal dynamics and laminar structure. Neuron, 98(2), 297-305.e6.

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Fluorescent Exosome Uptake Assay

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Purified exosomes were labeled with green fluorescent linker PKH-67 (Sigma) according to the manufacture’s protocol. Cells were seeded in 8-well chamber slides (8000 cells/well) and pre-treated with pharmacological inhibitors for 2 h. Then 5 μl of PKH67-dyed exosomes were added before a 4-h incubation to allow internalization. Finally, slides were washed twice with PBS, fixed with 4% paraformaldehyde (PFA), and mounted with DAPI-containing mounting media (Vector Labs). Images were taken using a Zeiss LSM 780 (Zeiss, Jena, Germany) confocal microscope.

Wei F., Ma C., Zhou T., Dong X., Luo Q., Geng L., Ding L., Zhang Y., Zhang L., Li N., Li Y, & Liu Y. (2017). Exosomes derived from gemcitabine-resistant cells transfer malignant phenotypic traits via delivery of miRNA-222-3p. Molecular Cancer, 16, 132.

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Immunofluorescence Staining of Kidney Proteins

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Immunofluorescence detection of proteins was performed as described previously [27 (link)]. Briefly, kidneys were perfused and fixed in 4% PFA for 2 h at room temperature, embedded in paraffin, and sectioned 8 μm thick. For immunofluorescence analysis of antibody stains, slides were washed in 1XPBS. For antigen retrieval, heat-induced epitope retrieval with Sodium Citrate buffer (pH 6.0) or Tris-EDTA (pH 9.0) was applied. To reduce non-specific binding, sections were blocked with normal serum (1:20). Primary and secondary antibodies were applied sequentially overnight at 4° C and 2 h at room temperature. Primary antibodies and dilutions were as follows: anti-TRPC6 antibody (1:100, ACC-120, Alomone Labs, Jerusalem, Israel), anti-P2Y2 antibody (1:100, APR-010, Alomone Labs, Jerusalem, Israel), anti-GFP antibody (GFP 1:200 Thermo Fisher Scientific, Waltham, MA). Alexa Fluor 488 and 594-conjugated secondary antibodies were purchased from Invitrogen (Waltham, MA). Slides were mounted by using DAPI-containing mounting media (VectaShield, Vector Laboratories Inc., Burlingame, CA). Sections were examined with Leica TCS SP8 (Leica Microsystems, Wetzlar, Germany) confocal/multiphoton laser scanning microscope systems as described previously [27 (link), 28 (link)].

GYARMATI G., TOMA I., IZUHARA A., BURFORD J.L., SHROFF U.N., PAPADOURI S., DEEPAK S, & PETI-PETERDI J. (2021). The role of TRPC6 calcium channels and P2 purinergic receptors in podocyte mechanical and metabolic sensing. Physiology international.

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Extracellular Vesicle Interactions in Insulin Secretion

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INS-1 cells were treated with varying amounts of PKH67-dyed (as per manufacturer’s protocol, Sigma-Aldrich) PC-Exo or PC-Exo with 10 nM AM inhibitor (AM 22–52, AnaSpec, Inc) for 24 hours. Cells were washed with 1x PBS and fixed with 4% paraformaldehyde. Duolink in situ kits were purchased from Olink Biosciences. Antibodies for AM (Phoenix Pharmaceuticals) and CRLR (Santa Cruz Biotechnology, Inc) were used for assessing AM/ADMR interactions. Antibodies for Bip (Abcam) and pro-insulin (Novus Biologicals) were used to detect Bip/pro-insulin in the presence of increasing PC-Exo. Conjugation of proximity ligation assay (PLA) probes, rolling circle amplification, and detection procedures were done according to the manufacturer’s instructions. Cells were mounted using DAPI-containing mounting media (Vector Labs). Images were taken on a Zeiss LSM 780 confocal microscope.

Javeed N., Sagar G., Dutta S.K., Smyrk T.C., Lau J.S., Bhattacharya S., Truty M., Petersen G.M., Kaufman R.J., Chari S.T, & Mukhopadhyay D. (2014). Pancreatic Cancer-derived Exosomes Causes Paraneoplastic β-cell Dysfunction. Clinical cancer research : an official journal of the American Association for Cancer Research, 21(7), 1722-1733.

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Resorufin Staining for Amyloid-Beta Vascular Deposits

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For the specific detection of Aβ-positive vessels, resorufin staining was performed in paraffin-embedded mice brain sections following the protocol described by Han et al. [75 (link)]. Briefly, all samples were deparaffinized, washed 3 times in PBS, and permeabilized in PBS-0.2% Triton X-100 (PBST) for 30 min. Samples were then immersed in 1 mM resorufin (Sigma-Aldrich) dissolved in PBST for 5 min. After rinsing with PBS, samples were rinsed with PBS-50% ethanol for 3 min and dehydrated. Finally, DAPI- containing mounting media (Vector Laboratories) was used as contrast staining. Samples were digitized at 20× objective using the Pannoramic 250 scanner (3DHistech) and the number of positive vascular deposits were manually determined in the selected area. Data are expressed as the number of Aβ-positive deposits per area (pixels²). Images were captured using Case Viewer Software (3DHistech).

Marazuela P., Paez-Montserrat B., Bonaterra-Pastra A., Solé M, & Hernández-Guillamon M. (2022). Impact of Cerebral Amyloid Angiopathy in Two Transgenic Mouse Models of Cerebral β-Amyloidosis: A Neuropathological Study. International Journal of Molecular Sciences, 23(9), 4972.

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TUNEL Assay for Apoptosis Quantification

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Cells were seeded at a cell density of 2 × 10⁴ cells/well in 2-chamber slides where drugs in complete media were added for 24 h at 37 °C with 5% CO₂. Treated cells were fixed in 4% paraformaldehyde (PFA) (Sigma- Aldrich, St. Louis, MO, USA), permeabilized by 0.1% Triton X-100 (Sigma- Aldrich, St. Louis, MO, USA) and stained with TUNEL assay kit (Roche, Mannheim, Germany) accordingly to the manufacturer’s instructions before being mounted with DAPI containing mounting media (Vector Laboratories, Burlingame, CA, USA). Slides were visualized under the Olympus IX71 fluorescence microscope at 20× magnification. Images were captured using CellSens imaging software.

Poh H.M., Chiou Y.S., Chong Q.Y., Chen R.M., Rangappa K.S., Ma L., Zhu T., Kumar A.P., Pandey V., B.a.s.a.p.p.a., Lee S.C, & Lobie P.E. (2019). Inhibition of TFF3 Enhances Sensitivity—and Overcomes Acquired Resistance—to Doxorubicin in Estrogen Receptor-Positive Mammary Carcinoma. Cancers, 11(10), 1528.

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Immunohistochemical Labeling of Catecholaminergic Neurons

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Sections were permeabilized for 1 hour in a 4% donkey serum/PBS blocking buffer containing 0,3% Triton X-100 (MilliporeSigma) and incubated overnight at 4°C with the following primary antibodies diluted in a 1% Donkey serum/PBS buffer: rabbit anti-TH (Abcam, EP1532Y, ab137869, 1:2,000) and goat anti-HA (Genscript, A00168, 1:500). Following incubation with primary antibodies, tissues were washed with PBS 3 times for 10 minutes and incubated for 1.5 hours at room temperature with a combination of corresponding donkey anti-species IgG conjugated to AlexaFluor probe (Invitrogen, 1:400). Tissues were then washed with PBS and mounted in DAPI-containing mounting media (Vectashield). Illustrative images were acquired using a confocal microscope Leica SP8 at the BioImaging Center in Bordeaux.

Arotcarena M.L., Bourdenx M., Dutheil N., Thiolat M.L., Doudnikoff E., Dovero S., Ballabio A., Fernagut P.O., Meissner W.G., Bezard E, & Dehay B. (2019). Transcription factor EB overexpression prevents neurodegeneration in experimental synucleinopathies. JCI Insight, 4(16), e129719.

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Quantitative Pancreatic β-Cell Analysis

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Formalin-fixed pancreas tissues were embedded in paraffin. Sections were deparaffinized, rehydrated, and incubated with blocking solution as previously described (15 (link)). Sections were incubated overnight at 4°C with antibodies against insulin (Dako) and Ki67 (Millipore), followed by secondary antibodies conjugated to FITC and Cy3 (Jackson ImmunoResearch). DAPI-containing mounting media (Vector Laboratories) was added to coverslips. β-Cell mass analysis entailed assessing total pancreas and insulin-positive cell areas from five insulin-stained sections (5 µm) separated by 200 µm and measured by using Image-Pro software (Media Cybernetics). β-Cell mass (average β-cell fraction multiplied by pancreas weight) assessment was performed using Surveyor software (Objective Imaging) automated scanning with a Leica fluorescent microscope (Leica Microsystems). Cell proliferation was analyzed using costaining of Ki67 with insulin on tissue sections of control and transgenic mice. Apoptotic cells in insulin-positive cells were assessed using the ApopTag Red In Situ Apoptosis Detection kit (Millipore) on tissue sections of control and transgenic mice.

Alejandro E.U., Bozadjieva N., Blandino-Rosano M., Wasan M.A., Elghazi L., Vadrevu S., Satin L, & Bernal-Mizrachi E. (2017). Overexpression of Kinase-Dead mTOR Impairs Glucose Homeostasis by Regulating Insulin Secretion and Not β-Cell Mass. Diabetes, 66(8), 2150-2162.

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