Dmi8 microscope system

Manufactured by Leica

Sourced in Germany

The Leica DMi8 is a high-performance microscope system designed for advanced imaging and analysis. It features a modular design that allows for customization and integration of various accessories and imaging techniques. The DMi8 provides reliable and precise performance for a wide range of applications.

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Lab products found in correlation

Bovine serum albumin (bsa), by Merck Group (2 mentions) Giemsa staining, by Merck Group (1 mentions) Normal goat serum (ngs), by Merck Group (1 mentions) Tween 20, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Las x, by Leica (1 mentions) Vectashield, by Vector Laboratories (1 mentions)

Spelling variants of this product

DMi8 microscope (744 citations) DMi8 system (10 citations) DMi8 inverted microscope system (9 citations) DMi8 fluorescence microscope system (4 citations) DMi8 M System fluorescence microscope (2 citations) Dmi8-S Inverted Microscope System (2 citations)

5 protocols using dmi8 microscope system

Karyotyping iPSCs via Giemsa Staining

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A Giemsa staining was performed to karyotype the iPSC culture. Briefly, medium of confluent iPSCs was replaced with medium containing 20 μL/mL KaryoMAX Colcemid solution and incubated for 1.5 hours in the incubator. Cells were then dissociated using TripLE Select for 7 minutes and centrifuged for 5 minutes at 1000 rpm. Cells were carefully resuspended in 1 mL 0.075 M prewarmed KCl, 2 mL extra KCl was added, and cells were incubated for 10 minutes in the incubator. Three drops of fixative (1:3 acetic acid: methanol) were added and suspension was centrifuged for 5 minutes at 1000 rpm. Cells were washed twice and resuspended in 20–30 drops fixative.
The suspension was dropped onto cold, moist microscope slides and left to dry completely. Giemsa staining (Sigma-Aldrich) was applied onto the slides for 5 minutes, washed in deionized water and imaged on a Leica DMi8 microscope system (Leica, Wetzlar, Germany) using the appropriate Leica Software (LAS X).

de Leeuw V.C., van Oostrom C.T., Imholz S., Piersma A.H., Hessel E.V, & Dollé M.E. (2020). Going Back and Forth: Episomal Vector Reprogramming of Peripheral Blood Mononuclear Cells to Induced Pluripotent Stem Cells and Subsequent Differentiation into Cardiomyocytes and Neuron-Astrocyte Co-cultures. Cellular Reprogramming, 22(6), 300-310.

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Immunofluorescence Labeling Protocol

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Samples were rinsed with PBS and fixed for 30 min with 4% prewarmed paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in PBS. Cells were permeabilised for 5 min with 0.2 % (Table 1, protocol A) or 0.5 % (protocol B) Triton X-100 (Sigma-Aldrich) in PBS. Blocking was performed either in 1% bovine serum albumin (BSA; w/v; Sigma-Aldrich), 0.5 % Tween-20 (Sigma-Aldrich) in PBS for 1 h at 37 • C (protocol A) or in 5% BSA (Sigma-Aldrich) for 30 min at RT(protocol B), depending on the antibody used (Table 1). Primary antibodies were applied overnight at 4 • C in 0.5 % BSA/0.5 % Tween-20 in PBS (protocol A) or 5% normal goat serum (v/v, Sigma-Aldrich)/0.5 % Tween-20 (Sigma-Aldrich) in PBS (protocol B). After washing away the primary antibodies, secondary antibodies were applied for 1 h in the same antibody incubation mixture. DAPI (Sigma-Aldrich) was used to stain nuclei of the cells. Imaging was performed on a Leica DMi8 microscope system (Leica, Wetzlar, Germany) using the appropriate Leica Software (LAS X). Images were further processed in ImageJ (version 1.51n; Rasband, 1997 Rasband, -2018 [25] [25] ).

de Leeuw V.C., van Oostrom C.T.M., Westerink R.H.S., Piersma A.H., Heusinkveld H.J, & Hessel E.V.S. (2020). An efficient neuron-astrocyte differentiation protocol from human embryonic stem cell-derived neural progenitors to assess chemical-induced developmental neurotoxicity. Reproductive toxicology (Elmsford, N.Y.), 98.

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Immunocytochemical Staining of iPSC and Cardiac Cells

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iPSC and neuronal differentiation cultures were stained as previously described by de Leeuw et al., 2019 (link). Cardiac differentiation cultures were stained using the Human Cardiomyocyte Immunocytochemistry Kit from Gibco. Primary and secondary antibodies used are listed in Table 2. Samples were imaged on a Leica DMi8 microscope system (Leica) using the appropriate Leica Software (LAS X). Image processing was performed in ImageJ (version 1.51n; Rasband, 1997–2018).

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Scratch Wound Healing Assay

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Cells were cultured in 6-well plates. After 24 h of incubation wounds were created by scraping the cells with a 200 μL pipette tip. Cells were then washed twice with phosphate-buffered saline (PBS) to clear away debris at the edge of the scratch and subsequently incubated in DMEM with 10% FBS. Microscopic images of the wounds at different time points were captured by the Leica DMi8 microscope system (Leica Microsystems, Wetzlar, Germany). The scratch assay was performed in triplicate.

Liu W., Yuan J., Liu Z., Zhang J, & Chang J. (2018). Label-Free Quantitative Proteomics Combined with Biological Validation Reveals Activation of Wnt/β-Catenin Pathway Contributing to Trastuzumab Resistance in Gastric Cancer. International Journal of Molecular Sciences, 19(7), 1981.

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Immunofluorescence Imaging of Nasal Epithelium

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Cryosections were prepared from nasal epithelium biopsies as described (19 (link)). Briefly, after PBS rinse and any required pretreatments, tissue sections were incubated in blocking solution with 5% donkey serum and 0.1% Triton X-100 for 30 minutes at room temperature. Primary antibodies (Supplemental Table 1) were diluted in the blocking solution and incubated overnight in a humidified chamber at 4°C. Detection by species-specific fluorescent-conjugated secondary antibodies (Jackson ImmunoResearch) was performed at room temperature for 45 minutes. Sections were counterstained with DAPI, and coverslips were mounted using Vectashield (Vector Labs) for imaging, using a Leica DMi8 microscope system (Leica Microsystems). Images were processed using ImageJ software, version 2.1.0/1.53c (NIH). Scale bars were applied directly from the Leica acquisition software metadata in ImageJ Tools.

Oliva A.D., Gupta R., Issa K., Abi Hachem R., Jang D.W., Wellford S.A., Moseman E.A., Matsunami H, & Goldstein B.J. (2022). Aging-related olfactory loss is associated with olfactory stem cell transcriptional alterations in humans. The Journal of Clinical Investigation, 132(4), e155506.

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