Oil Red O staining was performed according to the procedure described previously [51 (link)]. Briefly, C3H10T1/2 cells were treated with ADM containing 2 mg/mL of AE, ME, HE, CE, and WE or 1.5 mg/mL of BE and EE extracts for seven days. Cells were rinsed briefly with PBS, fixed, and air-dried before staining, then rinsed carefully with PBS, and viewed under an inverted microscope (Nikon Eclipse Ti–U inverted, Japan).
Eclipse ti u inverted
Manufactured by Nikon
Sourced in Japan
The Nikon Eclipse Ti-U is an inverted microscope designed for a variety of laboratory applications. It features a sturdy, ergonomic design and provides high-quality optical performance for observation and imaging. The core function of the Eclipse Ti-U is to enable detailed examination and analysis of specimens.
Automatically generated - may contain errors
Lab products found in correlation
Nis elements software, by Nikon (2 mentions)
Calcein am, by Thermo Fisher Scientific (2 mentions)
Nis elements f software, by Nikon (1 mentions)
N n dimethylformamide (dmf), by Merck Group (1 mentions)
Ds cooled camera head, by Nikon (1 mentions)
Donkey anti goat igg conjugated with alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Donkey anti rabbit igg conjugated with alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
C2 microscope, by Nikon (1 mentions)
Cryotome fse, by Thermo Fisher Scientific (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
Moflo xdp cell sorter, by Beckman Coulter (1 mentions)
11 protocols using eclipse ti u inverted
1
Oil Red O Staining of C3H10T1/2 Cells
2
Optogenetic Stimulation and Calcium Imaging
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The optogenetic stimulation and calcium imaging system was built around a modified Nikon Eclipse Ti-U inverted microscope that contained two stacked filter cube turrets. All imaging was conducted using a 10x, numerical aperture (NA) 0.45, Plano Apo objective. Worms were imaged on 6 cm NGM agarose plates on a Ludl BioPrecision2 XY motorized stage controlled by a MAC 6000 stage-controller. Custom real-time computer vision software kept the worm centered in the field of view via an automated feedback loop.
3
Scratch Wound Assay for A549 Cells
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A549 cells (2.0 × 105 cells/well) and A549-FLAG-BAP1 cells (2.0 × 105 cells/well) were seeded in 24-well plates and a straight wound was created by scratching with a yellow pipette tip. Cells were cultured in serum-free medium and allowed to migrate into the wound area and the images of cell movement were obtained using the Eclipse Ti-U inverted microscope (Nikon, Tokyo, Japan). Wounded areas were calculated using NIS Elements F software (Nikon, Tokyo, Japan).
4
Comparing Pantoea sp. YR343 and ΔcrtB Motility
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To compare the swimming motility function of Pantoea sp. YR343 and ΔcrtB cells, cells were grown overnight with shaking (250 rpm) in LB medium at 28 °C. Swimming motility was examined on LB containing 0.3% w/v agar. A 5 μL aliquot of cells were inoculated in the center of the plate and incubated at 28 °C for 18 h. Live cell imaging of bacterial motility was measured using a Nikon Eclipse Ti-U inverted microscope. Cells from motility plates were inoculated in R2A media overnight at 28 °C with shaking (250 rpm). Next day, cells were reinoculated in R2A media and grown to an OD600 of 0.5. A 20 µL aliquot of cells were placed on a coverslip and 10 s videos were captured using NIS-Elements imaging software. Trajectories and velocities (pixels/frame) of Pantoea sp. YR343 and ΔcrtB cells were calculated with the “TrackMate” plugin (https://imagej.net/TrackMate ).
5
Quantifying Intracellular Parasite Replication
6
3D Osteosarcoma Spheroid Culture
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A 3D cell culture is an improved method that can simulate the cell growth environment in vivo and is a more accurate model to determine the behavior of cancer cell growth. Following the manufacturer's protocol, osteosarcoma cell spheroid formation was established in HDP1096 Perfecta3D® 96-Well Hanging Drop Plates (3D Biomatrix). Hanging drops were formed by pipetting 40 μL of cell suspension (1 × 105/mL) into each well. Medium was changed every other day to provide enough nutrients for cells and to prevent osmolality shift of the medium. After seven days culture, the spheroids were harvested from the bottom side of the plate by gently pipetting 100 μL PBS into each well. After 15 minutes of incubation with 1 μM Calcein AM (Life Technologies), the spheroids were imaged on a Nikon Eclipse Ti-U inverted fluorescence microscope (Nikon Instruments, Inc NY, CA). The size of spheroids was calculated using ImageJ.
7
Osteoclast Differentiation and TRAP Staining
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BMMs were treated with M-CSF and RANKL for 3 days, and the generated osteoclasts were stained for TRAP by incubation in a solution containing fast red violet LB salt and naphthol AS-MX phosphate in N,N-dimethylformamide (Sigma-Aldrich, St. Louis, MO, USA). The solution was prepared according to the manufacturer’s instructions. Osteoclasts were first washed with phosphate buffered saline (PBS) and then fixed with 10% formalin for 5 min. After three washes with distilled water, the fixed cells were stained with fast red violet/naphthol AS-MX phosphate solution for 30–40 min and observed under a Nikon Eclipse Ti-U inverted microscope system (Nikon, Seoul, South Korea). Images of the stained cells were captured using a digital camera attached to the microscope. Multinuclear cells were manually counted.
8
Immunofluorescence Assay for F4+ ETEC/VTEC/EPEC
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Attachment of F4+ ETEC/VTEC/EPEC to ileal mucosa was determined by an indirect immunofluorescence assay. Ileal tissue samples fixed with 4% paraformaldehyde were embedded in paraffin, cut into 4-μm sections, and collected on silanized slides. The sections were then incubated with rabbit anti-F4 fimbriae antiserum (China Institute of Veterinary Drug Center, Beijing, China) overnight in a humidified chamber at 4 °C. Goat anti-rabbit Cy3-conjugated (AP307F; Sigma-Aldrich) was used as secondary antibody, and DAPI 3 (Sigma-Aldrich) was used for nuclear staining. The slides were visualized and photographed using a Nikon Eclipse Ti-U inverted fluorescence microscope equipped with a Nikon DS cooled camera head (Nikon).
9
Cryosectioning and Immunostaining of Mouse Limb Muscles
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The mouse limb muscles with bioluminescence signal were dissected and frozen in O.C.T compound (Sakura Finetek USA, Torrance, California) on dry ice and then placed into the −80°C freezer for a minimum of 24 hours. The tissue was then sectioned using a Cryotome FSE (Thermo Scientific, Waltham, Massachusetts) set to 10–15 μm. Immunostaining was performed as we previously described.35 The goat anti‐GFP antibodies or rabbit anti‐GFP antibodies (Novus Biologicals, Littleton, Colorado), rabbit anti‐human CD31 antibodies (Bethyl Laboratories, Montgomery, Texas), and sheep anti‐FVIII antibodies (Affinity Biologicals, Ancaster, Canada) were diluted in PBS with 1% bovine serum albumin (BSA) to 100, 200, and 50 folds, respectively. Secondary antibodies including donkey anti‐goat IgG conjugated with AlexaFluor 488 (ThermoFisher Scientific), donkey anti‐rabbit IgG conjugated with AlexaFluor 594 (ThermoFisher Scientific), and donkey anti‐sheep IgG conjugated with AlexaFluor 647 (EMD Millipore, Burlington, Massachusetts) were diluted to 500 folds in PBS with 1% BSA, respectively. Fluorescence images were captured using a Nikon Eclipse Ti‐U Inverted or Nikon C2 microscope (Nikon Instruments Inc, Melville, New York).
10
Quantifying Cellular Nitric Oxide Levels
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Cells
were seeded at a
density of 4 × 105 per well in 60 mm culture dishes.
After receiving treatment, cells were then washed in HBSS and incubated
in HBSS containing 5 μM 3-amino-4-aminomethyl-2′,7′-difluorescein
(DAF-FM-DA) for 20 min at 37 °C under 5% CO2. Cells
were imaged using a Nikon Eclipse Ti–U inverted microscope
(Nikon, Tokyo, Japan), and images were analyzed with the NIS elements
software (Nikon). In addition, DAF-FM-DA-stained cells were also analyzed
by a MoFlo XDP Cell Sorter (Beckman Coulter, Indianapolis, IN, USA)
for quantification of cellular NO levels.
were seeded at a
density of 4 × 105 per well in 60 mm culture dishes.
After receiving treatment, cells were then washed in HBSS and incubated
in HBSS containing 5 μM 3-amino-4-aminomethyl-2′,7′-difluorescein
(DAF-FM-DA) for 20 min at 37 °C under 5% CO2. Cells
were imaged using a Nikon Eclipse Ti–U inverted microscope
(Nikon, Tokyo, Japan), and images were analyzed with the NIS elements
software (Nikon). In addition, DAF-FM-DA-stained cells were also analyzed
by a MoFlo XDP Cell Sorter (Beckman Coulter, Indianapolis, IN, USA)
for quantification of cellular NO levels.
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