Leica dmr fluorescent microscope

Manufactured by Leica Microsystems

Sourced in Germany

The Leica DMR fluorescent microscope is a high-performance laboratory instrument designed for advanced imaging and analysis. The microscope utilizes fluorescence illumination to visualize and study fluorescently labeled samples. It is equipped with a range of optics and accessories to enable researchers to conduct detailed observations and experiments.

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Lab products found in correlation

4 6 diamidino 2 phenylindole dapi mounting medium, by Vector Laboratories (1 mentions) Wheat germ agglutinin (wga), by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions) Fluorometric tunel assay kit, by Promega (1 mentions) Baclight kit, by Thermo Fisher Scientific (1 mentions) Dibutylphthalate polystyrene xylene (dpx), by Electron Microscopy Sciences (1 mentions) Alexa fluor 488 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Donkey anti goat 488, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 donkey anti rabbit, by Thermo Fisher Scientific (1 mentions) Polyclonal rabbit anti th, by Merck Group (1 mentions) Superfrost plus microscope slide, by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Leica tcs sp8 confocal microscope, by Leica Microsystems (1 mentions)

5 protocols using leica dmr fluorescent microscope

Epithelial Cell Identification Protocol

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As in our previous studies [8 (link),49 (link)], epithelial cell origin was confirmed by the examination of cell morphology, the immunoflourescent staining of fixed cells for pancytokeratin, as well as standard diagnostic cytology and immunohistochemistry examination.
Ascitic culture cells were seeded onto coverslips, fixed, and hydrated. Coverslips were blocked with potassium chloride (KCl) buffer (120 mmol/L of KCl, 20 mmol/L of NaCl, 10 mmol/L of Tris-HCl, 1 mmol/L of ethylenediaminetetraacetic acid EDTA plus 0.1% Triton X-100) containing 2% (w/v) bovine serum albumin (BSA), and then incubated with 1:100 fluorescein isothiocyanate FITC-conjugated antipancytokeratin antibody (Merck Millipore, Watford, UK) for 1 h. Coverslips were mounted onto slides with 4´,6-diamidino-2-phenylindole (DAPI) mounting medium (Vectashield, Peterborough, UK). Cultures were examined with a Leica DMR fluorescent microscope (Leica microsystems GmbH, Wetzlar, Germany) and considered epithelial in origin if they contained >95% cytokeratin-positive cells.

Gentles L., Goranov B., Matheson E., Herriott A., Kaufmann A., Hall S., Mukhopadhyay A., Drew Y., Curtin N.J, & O’Donnell R.L. (2019). Exploring the Frequency of Homologous Recombination DNA Repair Dysfunction in Multiple Cancer Types. Cancers, 11(3), 354.

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Viability and Biofilm Formation of Microorganisms on Titanium

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After seeding fungal and bacterial cells on titanium discs (as described above), discs containing microorganisms were collected every 4 h for a period of 24 h, in triplicate. Plates were washed twice with PBS. Cell viability was measured by adding 300 µL of 1:10 v/v Presto Blue™ as described above. In addition, biofilm formation at different time points was monitored by fluorescence microscopy. Briefly, biofilms formed on titanium were stained in the dark in the following order: at 37 °C for 30 min with 25 µg/mL concavalin A–Alexa Fluor^® 488 conjugate (Molecular Probes, Eugene, OR, USA) to label the fungal cell wall; at 37 °C for 20 min with 5 µg/mL wheat germ agglutinin, Texas Red™-X Conjugate (WGA, Molecular Probes) to label the bacterial cell wall (although a caveat here is that WGA can also bind to chitin in the fungal cell wall); and at 37 °C for 10 min with 300 nM 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI, Molecular Probes) to label nucleic acids. These samples were visualized using a 63× objective lens in a Leica DMR fluorescent microscope (Leica Microsystems, Buffalo Grove, IL, USA).

Montelongo-Jauregui D., Srinivasan A., Ramasubramanian A.K, & Lopez-Ribot J.L. (2018). An In Vitro Model for Candida albicans–Streptococcus gordonii Biofilms on Titanium Surfaces. Journal of Fungi, 4(2), 66.

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Quantifying Apoptosis in Retinal Sections

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Apoptosis in retinas was analyzed using a fluorometric TUNEL assay kit as described previously (Promega) [31 (link)]. The 10 µm sections were used to measure the apoptotic cells. Apoptotic nuclei were labeled with fluorescein conjugated dUTP according to the manufacturer’s instructions. Using a Leica DMR fluorescent microscope (Leica Microsystems, Buffalo Grove, IL), TUNEL counts were collected from a 200 µm wide region of the superior and inferior retina at 200 µm intervals, beginning 200 µm from the optic disc (OD).
TUNEL counts were collected from at least three retinas from separate animals for each genotype for 1- and 3- month-old animals. The results were plotted as a spider plot with distance from the OD as the x-axis and TUNEL-positive nuclei per 0.01 mm² (area of 200 µm retina length × 50 µm average thickness) as the y-axis. Points located 0.4 mm to 1.2 mm away from the OD were considered central to the OD, while points located 1.6 mm and farther were considered peripheral. Plot points were analyzed for statistical significance using two-way analysis of variance (ANOVA) with Tukey’s correction for repeated measures (Prism, GraphPad, La Jolla, CA) [28 (link)].

Pak J.S., Lee E.J, & Craft C.M. (2015). The retinal phenotype of Grk1−/− is compromised by a Crb1rd8 mutation. Molecular Vision, 21, 1281-1294.

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Visualization of Bacterial Biofilm Viability

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The qualitative analysis of the CFS-treated and untreated biofilms developed by the different bacterial species was performed using live/dead staining (BacLight kit; Thermo Fisher Scientific, Waltham, MA, United States) as indicated by the manufacturers and fluorescent microscopy evaluation with a Leica DMR Fluorescent Microscope (Leica Microsystem, Wetzlar, Germany). Two experiments were performed in triplicate.

Vitale I., Spano M., Puca V., Carradori S., Cesa S., Marinacci B., Sisto F., Roos S., Grompone G, & Grande R. (2023). Antibiofilm activity and NMR-based metabolomic characterization of cell-free supernatant of Limosilactobacillus reuteri DSM 17938. Frontiers in Microbiology, 14, 1128275.

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Immunohistochemical Visualization of Neuronal Pathways

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Sections were rinsed in PB and incubated overnight at room temperature in a primary antibody solution. The primary antibody solution consisted of polyclonal rabbit anti-TH (Millipore, 1:1500), 3% normal donkey solution (Millipore) and 0.4% Triton X-100 (Sigma-Aldrich) in 0.1M PB. For retrograde tracing we also used goat anti-CTB (List Biological Solutions, 1:10,000). After incubation in the primary antibody, sections were washed and then incubated in a secondary antibody for two hours at room temperature. We used donkey anti-rabbit Alexa Fluor 568, and donkey anti-goat 488 to visualize the primary antibodies (1:250, Life Technologies). For anterograde experiments we used Alexa Fluor 488 conjugated streptavidin (1:250, Life Technologies) to visualize BDA. The sections were then mounted on Superfrost Plus microscope slides (Fisher Scientific), dehydrated and cleared, and coverslipped with DPX (Electron Microscopy Sciences). Label was observed using a Leica TCS SP8 confocal microscope or Leica DMR fluorescent microscope (Leica Microsystems).

Nevue A.A., Felix RA I.I, & Portfors C.V. (2016). Dopaminergic projections of the subparafascicular thalamic nucleus to the auditory brainstem. Hearing research, 341, 202-209.

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