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Dm6000 microscope

Manufactured by Leica camera
Sourced in Germany, United States, France

The Leica DM6000 is a high-performance microscope designed for advanced scientific research and analysis. It features a modular design, allowing for customization to meet specific experimental requirements. The DM6000 offers exceptional optical quality, delivering clear and detailed images. Its advanced technology enables precise control and data capture for accurate study and documentation of samples.

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83 protocols using dm6000 microscope

1

Histological Analysis of Mammary Tumors

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Mammary tumors, lung and liver tissues were fixed in 10% buffered formalin at room temperature for 48 h. Subsequently, the samples were embedded in paraffin and sectioned longitudinally at 5 μm. The tumor sections were stained with hematoxylin & eosin (H&E) and examined using a Leica DM6000 microscope. Images were taken using an Olympus BX53 F microscope (Tokyo, Japan) and analyzed with the Leica Application Suite (LAS) digital image processing software (Leica).
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2

Quantitative Immunofluorescence Analysis of Pancreatic Islet Cells

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Slides were dewaxed prior to heat induced antigen retrieval with Biocare Rodent Decloaker at 100 °C for 15 min. After cooling for 30 min the slides were blocked for 1 h with goat serum block. Slides were incubated overnight at 4 °C with 1:400 dilutions of both Invitrogen guinea pig anti-insulin and Cell Signaling’s rabbit anti-glucagon antibodies. After 17 h, the slides were washed in TBST prior to incubation with 1:300 dilutions of Invitrogen’s goat anti-guinea pig Alexa 488 and Invitrogen’s goat anti-rabbit Alexa 594 secondary antibodies for 1 h at RT. The slides were then washed in TBST prior to counterstaining with Hoechst.
After mounting and allowing the slides to cure for about 1–2 h, the slides were imaged on a Leica DM6000 microscope. Images were collected using a Hamamatsu EM-CCD (512 × 512 pixel) camera and were exported as single channel TIFF images for analysis in CellProfiler software. A CellProfiler pipeline was developed to identify insulin-positive beta cells as well as glucagon-positive alpha-cells based on green or red staining intensity, respectively. The area and integrated intensity of each cell type was collected and used to determine the average intensity of green (insulin) or red (glucagon) signal as well as the ratio of glucagon expressing vs insulin expressing cell area.
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3

Mitotic Cell Quantification Protocol

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Cells were fixed in 70% ethanol, washed 3 times with PBS and stained with DAPI before being visualized using the Leica DM6000 microscope. Cells were scored as mitotic when they were binucleates with no septum.
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4

Apoptosis Detection in SH-SY5Y Cells

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The In Situ Cell Death Detection Kit, POD (TUNEL assay, Roche Diagnostics GmbH, Mannheim, Germany), was used to assess apoptotic cells. SH-SY5Y cells were grown on coverslips. They were treated with 10 µM of suramin for 1 h before a 20 µM 6-OHDA incubation for 8 h. Cells were fixed for 1 h with a 4% paraformaldehyde solution after washing the cells thrice with PBS. Then, we blocked the cells with a 3% H2O2 solution in methanol for 10 min. The cells were incubated with a permeabilization solution for 2 min and washed thrice with PBS. Then, the TUNEL stain mixture was added, and the cells were incubated for 1 h at 37°C. Following this, 4′, 6-diamidino-2-phenylindole was added for 10 min, and the cells were washed thrice with PBS. Coverslips were mounted using glass slides and imaged using a Leica DM6000 microscope.
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5

Retinal Thickness and Length Measurements

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Measurements of ONL thickness and OS length were performed in the superior and inferior retina, located equidistant from the optic nerve head (ONH). Retina sections were analyzed by using a Leica DM-6000 microscope, with the objective Leica ∞/0.17/D, HCX PL FLUOTAR, 40X/0.75 that has an area of 0.31 mm2.
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6

In planta Transient Expression Visualization

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For visualization of in planta transient expression, confocal Laser Scanning Microscopy (CLSM) was done essentially as previously described (Laursen et al., 2016). Briefly, leaf discs from infiltrated tissue described above were excised, mounted in water, and observed by CLSM. Cell imaging was performed using an SP5x laser scanning confocal microscope equipped with a DM6000 microscope (Leica, Germany). Images were recorded using a 63x water immersion objective lens. Excitation/emission wavelengths were 488/500–550 nm for eGFP. The images were sequentially acquired and processed using the LAS X software (Leica).
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7

Fluorescent Protein Analysis with DAPI Staining

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Fluorescently tagged proteins and DAPI (4′, 6-diamidino-2-phenylindole) staining for nuclear analysis were visualized as described in [74 (link)]. A DM6000 microscope (Leica) equipped with a 100×/1.40 NA (numerical aperture) oil immersion objective and a DFC350 FX digital charge-coupled device camera (Leica) was used to image the cells. The obtained images were processed and analyzed with LAS AF (Leica) and ImageJ (http://rsbweb.nih.gov/ij/) software.
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8

Retinal Detachment Model in Mice

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Experimental neurosensory retinal detachment was induced in mice as described previously7 (link). Briefly, animals were anesthetized and a sclerotomy was performed using a 25G micro-vitrealretinal blade. A 35G beveled cannula (World Precision Instruments; Sarasota, FL, USA, Cat# NF35BV-2), attached to a NanoFil-100 syringe (World Precision Instruments, Cat# NANOFIL-100) was used to inject 2–3 µL of Healon (1% Hyaluronic Acid) (Abbott Medical Optics; Santa Ana, CA, USA, Cat# 05047450842) between the photoreceptor and RPE layers. Care was taken to detach approximately half of the retina in each animal. Only the sclerotomy was performed on the fellow eye as a control. After 3 days, animals were sacrificed and whole eyes were extracted and embedded in paraffin for sectioning as described above. TUNEL staining was performed using the DeadEnd™ Fluorometric TUNEL System (Promega Cat# G3250) and sections were counterstained using ProLong Gold Mountant with DAPI. Stained sections were imaged with a Leica DM6000 microscope using a 40X objective. TUNEL positive cells were manually counted across the detached portion of the retina. Counts were normalized to the total number of nuclei in the outer nuclear layer in the detached region, counted manually or with an automated cell counting macro using ImageJ7 (link),56 (link).
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9

Immunofluorescence Microscopy for Tubulin

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Immunofluorescence was carried out as described in [35 (link)]. Anti-tubulin (Abcam) and anti–rat FITC (Jackson ImmunoResearch) antibodies were used at 1:250. Microscope preparations were imaged using a DM6000 microscope (Leica) equipped with a 100x/1.40 NA oil immersion objective lens, A4, L5, and TX2 filters, and a DF350 digital charge-coupled device camera (Leica). Pictures were processed with LAS AF (Leica) and ImageJ (http://rsbweb.nih.gov/ij/) software.
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10

3D Imaging of Cellular Responses to NCS

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Three-dimensional stacks were captured every 10 min for a total of 320 min upon NCS addition using the Leica DM6000 microscope with Leica CSU22 spinning disc and Andor Ixon 897 camera. Twenty different cells were imaged for each condition (±NCS).
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