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Dm6000 microscope

Manufactured by Leica
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Sourced in Germany, United States, France, Switzerland, United Kingdom

The Leica DM6000 is a high-performance microscope designed for advanced research and analysis. It features a modular design that allows for the integration of various illumination and imaging technologies to suit a wide range of applications. The DM6000 offers exceptional optical performance, delivering clear and detailed images for a variety of sample types.

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Lab products found in correlation

Dfc390 camera, by Leica (5 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (4 mentions) Lsm 880 confocal microscope, by Zeiss (4 mentions) Coolsnap hq ccd camera, by Teledyne (4 mentions) High sensitivity 3ccd video camera system, by MBF Biosciences (4 mentions) Matlab, by MathWorks (4 mentions) In situ cell death detection kit, by Roche (3 mentions) Las af, by Leica (3 mentions) Donkey serum, by Merck Group (3 mentions) Stereo investigator software, by MBF Biosciences (3 mentions) Application suite v4 program, by Leica (3 mentions) Fv1000, by Olympus (3 mentions) Csu22 spinning disc, by Oxford Instruments (2 mentions) El6000 metal halide lamp, by Leica (2 mentions) Dfc350 fx monochrome ccd camera, by Leica (2 mentions) Goat anti glucagon α cell, by Cell Signaling Technology (2 mentions) Glass slide, by Thermo Fisher Scientific (2 mentions) Normal donkey serum (nds), by Merck Group (2 mentions) Smi 32p 100, by R&D Systems (2 mentions) Af1387, by R&D Systems (2 mentions)

158 protocols using dm6000 microscope

1

Quantifying Retinal Cell Apoptosis by TUNEL

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The number of apoptotic cells was analyzed by TdT-mediated dUTP nick end labeling (TUNEL), using the In Situ Cell Death Detection Kit, Fluorescein (Roche 11684795910) following the manufacturer’s directions. Twelve-micrometer cryosections were collected on slides and submit to TUNEL assay. To consider the presence of unspecific results, some retina sections were incubated with the reaction mix without TUNEL reaction enzyme. Sections were observed with a Leica DM-6000 microscope and then confocal images were acquired using the LSM710 Zeiss Confocal Microscopy system. The number of TUNEL-positive cells was evaluated in the central part of the retina by manual counts with 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.
Intartaglia D., Giamundo G., Marrocco E., Maffia V., Salierno F.G., Nusco E., Fraldi A., Conte I, & Sorrentino N.C. (2020). Retinal Degeneration in MPS-IIIA Mouse Model. Frontiers in Cell and Developmental Biology, 8, 132.
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2

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).
Chen C., Nong Z., Xie Q., He J., Cai W., Tang X., Chen X., Huang R, & Gao Y. (2017). 2-Dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione inhibits the growth and metastasis of breast carcinoma in mice. Scientific Reports, 7, 6704.
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3

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.
Hegde V., Na H.N., Dubuisson O., Burke S.J., Collier J.J., Burk D., Mendoza T, & Dhurandhar N.V. (2015). An adenovirus-derived protein: A novel candidate for anti-diabetic drug development. Biochimie, 121, 140-150.
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4

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.
Anda S., Boye E, & Grallert B. (2014). Cell-Cycle Analyses Using Thymidine Analogues in Fission Yeast. PLoS ONE, 9(2), e88629.
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5

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.
Feng C.W., Chen N.F., Chan T.F, & Chen W.F. (2020). Therapeutic Role of Protein Tyrosine Phosphatase 1B in Parkinson's Disease via Antineuroinflammation and Neuroprotection In Vitro and In Vivo. Parkinson's Disease, 2020, 8814236.
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6

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.
Intartaglia D., Giamundo G., Naso F., Nusco E., Di Giulio S., Salierno F.G., Polishchuk E, & Conte I. (2022). Induction of Autophagy Promotes Clearance of RHOP23H Aggregates and Protects From Retinal Degeneration. Frontiers in Aging Neuroscience, 14, 878958.
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7

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).
Almeida A., Dong L., Thorsen T.H., Raadam M.H., Khakimov B., Carreno-Quintero N., Kampranis S.C, & Bak S. (2022). Metabolic engineering of cucurbitacins in Cucurbita pepo hairy roots. Frontiers in Plant Science, 13, 1021907.
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8

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.
de Oya I.G., Manzano-López J., Álvarez-Llamas A., Vázquez-Aroca M.D., Cepeda-García C, & Monje-Casas F. (2023). Characterization of a novel interaction of the Nup159 nucleoporin with asymmetrically localized spindle pole body proteins and its link with autophagy. PLOS Biology, 21(8), e3002224.
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9

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).
Weh E., Lutrzykowska Z., Smith A., Hager H., Pawar M., Wubben T.J, & Besirli C.G. (2020). Hexokinase 2 is dispensable for photoreceptor development but is required for survival during aging and outer retinal stress. Cell Death & Disease, 11(6), 422.
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10

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.
Muñoz-Barrera M., Aguilar I, & Monje-Casas F. (2015). Dispensability of the SAC Depends on the Time Window Required by Aurora B to Ensure Chromosome Biorientation. PLoS ONE, 10(12), e0144972.
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