Dmi6000b

Manufactured by Leica

Sourced in Germany, United States, Switzerland, United Kingdom, Japan, Canada, France, China

The DMI6000B is a high-performance inverted research microscope designed for advanced microscopy applications. It features a modular design and a wide range of accessories to support various imaging techniques, such as brightfield, phase contrast, and fluorescence microscopy.

Automatically generated - may contain errors

Lab products found in correlation

Las af software, by Leica (25 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (20 mentions) Dfc365fx, by Leica (17 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (15 mentions) Alexa fluor 488, by Thermo Fisher Scientific (13 mentions) Metamorph software, by Molecular Devices (12 mentions) Las x software, by Leica (9 mentions) Orca r2, by Hamamatsu Photonics (9 mentions) Matrigel, by BD (9 mentions) El6000, by Leica (9 mentions) Hoechst 33342, by Thermo Fisher Scientific (8 mentions) Tcs sp5, by Leica (7 mentions) Triton x 100, by Merck Group (7 mentions) Application suite software, by Leica (7 mentions) Paraformaldehyde (pfa), by Merck Group (7 mentions) Dfc350fx, by Leica (6 mentions) Dfc360fx ccd camera, by Leica (6 mentions) Las af lite, by Leica (6 mentions) Lsm 710, by Zeiss (6 mentions) Lsm 880, by Zeiss (6 mentions)

1 065 protocols using dmi6000b

Neonatal Rat Cardiomyocyte and AC16 Cell Imaging

Check if the same lab product or an alternative is used in the 5 most similar protocols

Neonatal rat cardiomyocytes were stained with alpha-sarcomeric actinin (#MA1-22863 Thermo Scientific, USA) and DAPI. AC16 cells were stained with wheat germ agglutinin (#MP00831 Thermo Scientific, USA) and DAPI. All images were taken with 20X objective using Leica DMI 6000 B (Leica Microsystems, USA). Cell size was calculated with ImageJ software. All the experiments were repeated three times (biological replicate) with three wells for each group (technical replicate). More than a hundred cells were counted from each well.
(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted March 2, 2023. ; https://doi.org/10.1101/2023.03.01.530580 doi: bioRxiv preprint Paraffin-embedded heart sections were deparaffinized and stained with wheat germ agglutinin (#MP00831 Thermo Scientific, USA) and DAPI. Images from different regions of the heart were taken with a 20X objective using Leica DMI 6000 B (Leica Microsystems, USA). Cell size was measured using ImageJ software.

Singh S., Gaur A., Sharma R.K., Kumari R., Prakash S., Kumari S., Chaudhary A.D., Prasun P., Pant P., Hunkler H., Thum T., Jagavelu K., Bharati P., Hanif K., Chitkara P., Kumar S., Mitra K, & Gupta S.K. (2023). Musashi-2 causes cardiac hypertrophy and heart failure by inducing mitochondrial dysfunction through destabilizing Cluh and Smyd1 mRNA. Basic research in cardiology, 118(1).

+ Open protocol

+ Expand

Microscopic Analysis of Root Mucilage

Check if the same lab product or an alternative is used in the 5 most similar protocols

Roots were mounted on a microscope slide in a drop of water and examined directly for morphological analyses using a bright-field microscope (Leica DMI6000B, Wetzlar, Germany) and a stereomicroscope (Leica M125, Wetzlar, Germany). To examine the mucilage around BCs, India ink (Salis International Inc., Oceanside, CA, USA) staining was performed following a procedure adapted from Miyasaka and Hawes [33 (link)]. Briefly, mucilage was collected by dipping roots in a drop of water on a microscopic slide, and India ink was added by capillarity action between the slide and the cover slip. Root mucilage was observed using a bright-field microscope (Leica DMI6000B, Wetzlar, Germany). Mucilage from 30–35 roots was observed to ensure representativity for each set of observations.

Ropitaux M., Bernard S., Schapman D., Follet-Gueye M.L., Vicré M., Boulogne I, & Driouich A. (2020). Root Border Cells and Mucilage Secretions of Soybean, Glycine Max (Merr) L.: Characterization and Role in Interactions with the Oomycete Phytophthora Parasitica. Cells, 9(10), 2215.

+ Open protocol

+ Expand

Analysis of Germ Cell Differentiation in C. elegans

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Synchronized L4-staged DUP17, DUP36 (non-green pharynx), and DUP49 worms were live mounted on agarose slides. Images of the bend in the gonad were acquired for each strain using fixed exposure conditions. 60× objective on a Leica DMI6000B – 10 gonad arms/strain.
To count sperm, synchronized young adults were fixed in M9 with 8% PFA for 1 h, washed 3× with PBS, 1× with 95% ethanol for 1 min, 3× with PBS then mounted on a charged slide with mounting media containing DAPI. Sperm nuclei were imaged and counted in each spermatheca using a 60× objective on a Leica DMI6000B – 10 worms/strain.
To count brood size, six L4 stage worms were picked to a plate and passaged to new plates each day for six days. The number of progeny were totaled from each plate and divided by six. This was repeated three times to get the average broods of N2 versus pqn-75(tm6575) and N2 fed control (empty vector) versus pqn-75 RNAi. RNAi feeding was performed as previously described (Kamath et al., 2001 (link)).

Rochester J.D., Tanner P.C., Sharp C.S., Andralojc K.M, & Updike D.L. (2017). PQN-75 is expressed in the pharyngeal gland cells of Caenorhabditis elegans and is dispensable for germline development. Biology Open, 6(9), 1355-1363.

+ Open protocol

+ Expand

Adipogenic Induction of ASC Spheroids

Check if the same lab product or an alternative is used in the 5 most similar protocols

At week 5 of adipogenic induction, ASC spheroids from sSAT, sRC, and dSAT were washed in PBS and were transferred to 4% paraformaldehyde in PBS at room temperature for 1 h. Subsequently, induced ASC spheroids were washed in PBS and placed in successive baths of 15% and 30% sucrose solution at room temperature for 24 h each. The ASC spheroids were embedded in optimal cutting temperature (OCT, Tissue-Tek) and maintained in a −80°C freezer until sectioning. Sections of 10 μm were obtained using a cryostat (Leica DMI 6000 B) and subsequently collected onto 0.01% poly-L-lysine-coated slides (Sigma). The slides with the sections were stored in a freezer at −20°C until staining. Cryosections were left at room temperature for 15 min and stained with 1 mg/mL Nile red (Sigma) diluted (1:50) in PBS. The cell nucleus was stained with 0.5 μg/mL Hoechst. The images were obtained with the aid of a fluorescence microscope (Leica DMI 6000B) (Mannheim, Germany) with LAS AF software (Leica, Mannheim, Germany). The laser microscope was programmed to stimulate at a range of 640–720 nm. Two independent analyses were evaluated, with a total of 324 spheroids of each sample obtained from four independent experiments.

Côrtes I., Alves G., Claudio-Da-Silva C, & Baptista L.S. (2023). Mimicking lipolytic, adipogenic, and secretory capacities of human subcutaneous adipose tissue by spheroids from distinct subpopulations of adipose stromal/stem cells. Frontiers in Cell and Developmental Biology, 11, 1219218.

+ Open protocol

+ Expand

Fluorescence-based Assays for Plant Cell Viability, Cellulose, and extracellular DNA

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell viability was assessed with fluorescein diacetate (FDA) (Sigma-Aldrich, Saint-Quentin Fallavier, France) as previously described by Jones and Senft [42 (link)]. Roots were incubated with the fluorescent probe (0.5 mg·L⁻¹) for 3 min and observed using an epifluorescence microscope (Leica DMI6000B, Wetzlar, Germany; λExcitation: 359 nm; λEmission: 461 nm).
Cytochemical staining of cellulose was carried out using Direct Red 23 (Sigma-Aldrich, Saint-Quentin Fallavier, France) as described previously [43 (link)]. Roots were incubated with the probe (0.1 mg·mL⁻¹) for 30 min in darkness. After 3 washes with distilled water, roots were observed using a confocal laser-scanning microscope (Leica TCS SP5, Wetzlar, Germany; λExcitation: 560 nm; λEmission: 570–655 nm).
Staining of exDNA was performed as described by Wen et al. [44 (link)]. Sytox Green (Invitrogen, Carlsbad, CA, USA) solution was made by 1:1000 dilution in sterile water. Mucilage was released from root tips by contact with the surface of the glass slide, and 10 μL of diluted Sytox Green were added to the sample, which was then covered with a cover slip and observed using an epifluorescence microscope (Leica DMI6000B, Wetzlar, Germany; λExcitation: 359 nm; λEmission: 461 nm).

+ Open protocol

+ Expand

Visualizing Adipocyte Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols

The differentiating adipocytes were observed by phase contrast imaging using a Leica DMI6000 B inverted microscope. Fatty acid visualization was conducted with Oil Red O (Sigma-Aldrich, Cat. No. O-0625) staining. To stain the cells, cells were washed twice with PBS, followed by fixation with a 10% formalin solution. After two washes with 60% isopropanol, cells were stained with 0.3% (w/v) Oil Red O in 60% isopropanol. Upon removal of Oil Red O, cells were washed twice with PBS and visualized using a Leica DMI6000 B inverted microscope.

Crown S.B., Marze N, & Antoniewicz M.R. (2015). Catabolism of Branched Chain Amino Acids Contributes Significantly to Synthesis of Odd-Chain and Even-Chain Fatty Acids in 3T3-L1 Adipocytes. PLoS ONE, 10(12), e0145850.

+ Open protocol

+ Expand

In Vitro Cell Invasion and Migration Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

For in vitro invasion and migration assays, SCC25 or CAL27 cells (50,000 cells/well) were plated into Transwell inserts (Corning, One Riverfront Plaza, NY, USA) that had been coated with Matrigel (BD Biosciences, Franklin Lakes, New Jersey, USA) or left uncoated. The lower chambers were filled with medium plus 20% FBS. After incubation at 37°C for 16 h, the penetrated cells were fixed with 4% paraformaldehyde (Solarbio, Beijing, China) and stained with 0.1% crystal violet (Solarbio). Each image was observed using an inverted microscope (DMI6000B, Leica, Germany).
A wound-healing assay was used to confirm the result of the Transwell assay. We added 500,000 SCC25 or CAL27 cells into six-well plates. When the cells had grown to approximately 80% confluence, we made scratches using 10-μL pipette tips, creating a wound field of approximately 400 mm wide, based on the scaleplate in the microscope. The plates were incubated in fresh medium (without FBS) for 48 hours. Images of gaps from several randomly selected fields were acquired at the start (0 h), halfway point (24 h), and endpoint (48 h) of the experiment, using an inverted microscope (DMI6000B, Leica).

Wang Y., Wang S., Wu Y., Ren Y., Li Z., Yao X., Zhang C., Ye N., Jing C., Dong J., Zhang K., Sun S., Zhao M., Guo W., Qu X., Qiao Y., Chen H., Kong L., Jin R., Wang X., Zhang L., Zhou J., Shen Q, & Zhou X. (2017). Suppression of the Growth and Invasion of Human Head and Neck Squamous Cell Carcinomas via Regulating STAT3 Signaling and miR-21/β-catenin Axis with HJC0152. Molecular cancer therapeutics, 16(4), 578-590.

+ Open protocol

+ Expand

Quantifying Continual Efferocytosis in Macrophages

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

After induction of apoptosis, Jurkat cells were stained with PKH26, PKH67 (Sigma-Aldrich), or pHrodo (Thermo Fisher) according to the manufacturer’s protocol. For single efferocytosis assays, macrophages were incubated with PKH26- or pHrodo- ACs for 45 minutes at a 5:1 AC:macrophage ratio. After 45 minutes, unbound ACs were removed by vigorous rinsing of the monolayer, and fluorescence and bright-field images were captured using an epifluorescence microscope (Leica DMI6000B) to identify the uptake of red-labeled ACs. For continual efferocytosis assays, macrophages were first incubated with PKH67-stained ACs (green) for 45 minutes at a 5:1 AC:macrophage ratio, followed by removal of unbound ACs and incubation in complete culture medium for 2 hours. The macrophages were then incubated with a second round of PKH26- or pHrodo-stained ACs (both red) for 45 minutes at a 5:1 AC:macrophage ratio. After 45 minutes, unbound ACs were removed, and fluorescence and bright-field images were captured using an epifluorescence microscope (Leica DMI6000B) with Leica LAS AF to quantify the number of macrophages with green plus red ACs as a percentage of macrophages with only green ACs, which is a measure of continual efferocytosis^{9 (link)}.

Danielsson O, & Jörnvall H. (1992). "Enzymogenesis": classical liver alcohol dehydrogenase origin from the glutathione-dependent formaldehyde dehydrogenase line.. Proceedings of the National Academy of Sciences of the United States of America, 89(19), 9247-9251.

+ Open protocol

+ Expand

Live-cell Microscopy for Cellular Dynamics

Check if the same lab product or an alternative is used in the 5 most similar protocols

Unless otherwise indicated, cells were imaged using a Quorum spinning disk microscope with a 10× or 20×, 1.0 NA objectives, or a 63×, 1.4 NA oil immersion objective (Leica DMI6000B inverted fluorescence microscope with a Yokogawa spinning disk head and Hamamatsu ORCA Flash4 sCMOS camera) and Volocity 6.3 acquisition software (Quorum). Confocal z-stacks of 0.3 μm were acquired. Images were analyzed with the Volocity software or Fiji v2.14.0 (ImageJ) and then imported and assembled in Adobe Illustrator v25.3.1 for labeling. For live cell imaging, cells were seeded in μ-Slide 8-well glass bottom chambers (ibidi). Twenty-four hours after seeding, growth media was replaced with live cell imaging media (RPMI with l-Glutamine and 25 mM HEPES (Wisent) supplemented with 10% FBS (Wisent)) containing the respective treatment condition. Cells were imaged at 37 °C using a Leica DMI 6000B inverted fluorescence microscope with a Yokogawa spinning disk head and Hamamatsu ImagEM X2 camera. Images were taken with a z-spacing of 0.5 μm. For invasion ruffle volume measurements, confocal z-stacks of 0.3 μm were acquired.

Zhu H., Sydor A.M., Boddy K.C., Coyaud E., Laurent E.M., Au A., Tan J.M., Yan B.R., Moffat J., Muise A.M., Yip C.M., Grinstein S., Raught B, & Brumell J.H. (2024). Salmonella exploits membrane reservoirs for invasion of host cells. Nature Communications, 15, 3120.

+ Open protocol

+ Expand

EV Internalization in Skin Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Next, in order to determine the internalization of MSC‐derived EVs by HSFs and HaCaT cells, 10 μg of EVs was labelled using green fluorescent dye (PKH67; Sigma) and incubated with 1 × 10⁵ cells at 37°C for 3 hours. The cells were then fixed with 4% paraformaldehyde for 15 minutes followed by the addition of 4',6‐diamidino‐2‐phenylindole (DAPI) (0.5 mg/mL; Invitrogen) for nucleus staining. Finally, green fluorescence was observed under a fluorescence microscope (Leica DMI6000B, Solms, Germany).
Next, to investigate the internalization of MSC‐derived EVs carrying miR‐27b by HSFs and HaCaT cells, Lipofectamine 3000 reagent (L3000001, Invitrogen) was employed to transfect the cy3‐conjugated miR‐27b (GenePharma, Shanghai, China) into MSCs in a serum‐free medium. After 6 hours, the medium was renewed using a medium containing 10% serum without EVs for further incubation for 48 hours. The cell supernatant was then collected, centrifuged and resuspended using PBS, which was then added into the HSFs and HaCaT cells, respectively. Finally, the cells were fixed with 4% paraformaldehyde for 15 minutes and added with DAPI (0.5 mg/mL; Invitrogen) for nucleus staining, followed by fluorescence microscopic observation (Leica DMI6000B, Solms, Germany).

Cheng S., Xi Z., Chen G., Liu K., Ma R, & Zhou C. (2020). Extracellular vesicle‐carried microRNA‐27b derived from mesenchymal stem cells accelerates cutaneous wound healing via E3 ubiquitin ligase ITCH. Journal of Cellular and Molecular Medicine, 24(19), 11254-11271.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!