Cells were plated at a concentration of 3 × 104 cells/cm2 and cultured in DMEM-low glucose (Sigma-Aldrich), 10% FBS (HyClone, Thermo Fisher Scientific), 4 mM L-glutamine (Euroclone), and 1% antibiotic-antimycotic mixture (Euroclone), with the addition of the mesenchymal stem cell adipogenesis kit (Millipore) for 21 days, according to the manufacturer's instructions. The adipogenic medium was changed every other day. At day 21, Oil Red O solution (Millipore) was used to stain lipid droplets of derived adipocytes, according to the manufacturer's procedures. All photomicrographs were acquired with an Axiovert 40 microscope (Zeiss) equipped with a Moticam 2300 camera (Motic). The mRNA expression of adipogenic markers including PPAR-γ and LPL were also assessed on days 7 and 21 by real-time PCR, as described above [17 (link)].
Axiovert 40 microscope
Manufactured by Zeiss
Sourced in Germany
The Axiovert 40 is an inverted microscope designed for routine observation and image capture. It features high-quality optics and illumination to provide clear, detailed images of specimens. The Axiovert 40 is suitable for a variety of applications, including cell culture monitoring, tissue analysis, and basic research.
Automatically generated - may contain errors
Lab products found in correlation
Moticam 2300 camera, by Motic (7 mentions)
Antibiotic antimycotic mixture, by Euroclone (6 mentions)
Dmem low glucose, by Merck Group (6 mentions)
L glutamine, by Euroclone (6 mentions)
Mesenchymal stem cell adipogenesis kit, by Merck Group (3 mentions)
Oil red o solution, by Merck Group (3 mentions)
Quant it picogreen, by Thermo Fisher Scientific (3 mentions)
Alizarin red solution, by Merck Group (3 mentions)
Axiocam erc 5s, by Zeiss (2 mentions)
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Easyscan 2 controller, by Nanosurf (2 mentions)
L ascorbic acid 2 phosphate, by Merck Group (2 mentions)
β glycerophosphate, by Merck Group (2 mentions)
Prolong gold antifade, by Thermo Fisher Scientific (2 mentions)
Axiocam hrc, by Zeiss (1 mentions)
S4500 microscope, by Hitachi (1 mentions)
Dimension 3100, by Bruker (1 mentions)
Axiovert mrm monochrome camera, by Zeiss (1 mentions)
Biotinylated anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
29 protocols using axiovert 40 microscope
1
Adipogenic Differentiation of Mesenchymal Stem Cells
2
Graphite to Graphene Oxide Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Graphite powder was purchased from Sinocarbon Materials Technology Co. GO was synthesized via the Hummers’ method (35 ) and exfoliated following the procedure previously described (30 (link)). GO was deposited directly on the BALM substrate from the water solution or using the so-called bubble deposition method for samples imaged in air (30 (link)). Au (5 nm) and Cr/Au (2/5 nm) near-ARA layers were received from WATCH LIVE SAS (Lyon, France) or fabricated in the laboratory. Trimethyl-(2-oxo-2-pyrene-1-yl-ethyl)-ammonium bromide was prepared according to the procedure by Nakashima et al. (34 ). GO and r-GO were imaged using a Zeiss Axiovert 40 microscope equipped with a 63× Plan Apo oil immersion objective (numerical aperture of 1.40) and a Zeiss AxioCam HRc. AFM images were performed using a Dimension 3100 from Bruker (Digital Instruments); SEM images were performed using a Hitachi S4500 microscope. Chronoamperometry was carried out using a VoltaLab PGZ 301 potentiostat, in a three-electrode electrochemical cell developed for BALM under atmospheric conditions. The gold near-ARA layer was used as working electrode, a gold wire was used as counter electrode, and the Ag/AgCl electrode was used as the reference electrode.
3
ESE-15-ol and 3MA Treatment Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Viable cells were seeded at a density of 375,000 cells/3 ml growth medium in 6 well plates. After attachment, cells were exposed to ESE-15-ol in the presence or absence of 3MA for 24 h (37 °C) along with appropriate controls. PlasDIC images were viewed at a 40× magnification with a Zeiss Axiovert-40 microscope (Göttingen, Germany) and captured with the Zeiss Axiovert MRm monochrome camera (Göttingen, Germany).
4
Retroperitoneal Adipose Tissue Immunohistochemistry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Retroperitoneal adipose tissue was included in Tissue-Tek® medium, frozen in liquid nitrogen and 5 μm sections were obtained and adhered onto silanized slides. Slides were stained with HE. Evaluation of leukocyte infiltrate was also performed by immunohistochemistry. Slides were blocked with 3% BSA and incubated overnight with mouse anti-rat CD11b/c mouse antibody (BD Pharmingen ™, 1:1000). Subsequently, the tissue was incubated with biotinylated anti-mouse IgG (Jackson ImmunoResearch, 1:200) secondary antibody for 2 hours, washed with 0.1M phosphate buffer and incubated with VectaStain ABC kit (Vector Laboratories, 1:100) for 2 hours. Detection of the antigen-antibody complex was performed through the chromogen 3,3'-diaminobenzidine (DAB) for 5 minutes at room temperature. Sections without the primary antibody (Cd11b/c) were used as negative control of the immunolabeling process. The qualitative evaluation of the slides was performed using photomicrographs captured by Zeiss Axiovert 40 microscope, with a 40x objective (Camera: Zeiss AxioCam ERc 5s).
5
Quantifying Neutrophil Extracellular Traps
6
Adipogenic Differentiation of hTSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hTSCs were plated at a concentration of 3 × 104 cells/cm2 in normal growth medium, and then switched to DMEM-low glucose (Sigma-Aldrich), 10 % FBS (HyClone, Thermo-Fisher Scientific), 4 mM L-glutamine (Euroclone), 1 % antibiotic-antimycotic mixture (Euroclone), with the addition of the mesenchymal stem cell adipogenesis kit (Millipore) for 21 days, according to the manufacturer’s instructions. At day 21, Oil Red O solution (Millipore) was used to stain lipid droplets of derived adipocytes, according to the manufacturer’s procedures. All photomicrographs were acquired with an Axiovert 40 microscope (Zeiss) equipped with a Moticam 2300 camera (Motic). The adipogenic medium was changed every 2–3 days.
7
Zonulin Localization in Caco-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A monolayer of Caco-2 cells was seeded onto glass cover slips in a 24-well plate at a density of 2 × 106 cells and grown to 50% to 60% confluence. The cells were then similarly exposed to Pseudomonas fluorescens as previously described. Subsequently, the cells were fixed in 1% paraformaldehyde, permeabilized with 0.2% Triton X-100, and washed in PBS. Mouse monoclonal antibody against human zonulin (1:50 dilution, Abcam, U.S.) was added to the cells and incubated for more than 12 hrs at 4 °C. After incubation with rhodamine-conjugated goat anti-mouse IgG (1:200 dilution) for 30 min, the cells were washed again with PBS. Nuclei were stained with 1 μg/mL DAPI for 3 min. The localization of zonulin in the cells was examined under an Axiovert 40 microscope (Carl Zeiss, Germany).
8
Chondrogenic Differentiation of hTSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hTSCs were maintained in a 3D culture by growing them in cell pellets (1 × 106 cells/pellet) in AdvanceSTEM chondrogenic differentiation medium (HyClone, Thermo Scientific), according to the manufacturer’s instructions. After 28 days of differentiation, matrix deposition by derived chondroblasts was detected with Alcian Blue staining (Sigma-Aldrich), according to the manufacturer’s instruction. All photomicrographs were acquired with an Axiovert 40 microscope (Zeiss) equipped with a Moticam 2300 camera (Motic). The chondrogenic medium was changed every 2–3 days.
9
Quantifying Neutrophil Extracellular Traps
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neutrophils were isolated using density gradient centrifugation. Cells were plated in a 96 well plate at approximately 1.5 × 104cells per well in Hank's Balanced Salt Solution (HBSS). Neutrophils were then stimulated with platelet activating factor (PAF, 0-50μm, Millipore, #511075) for 30 minutes. Cells were fixed with 3% paraformaldehyde and then DNA was stained with Hoechst 33342 (Molecular Probes, #H-3570). NETs were visualized using a Zeiss Axiovert 40 microscope under 10x-40xmagnification. Supernatant was collected, spun at 14g × 10 min and the level of DNA were measured using Quant-iT™Picogreen© (Invitrogen, MP07581), a fluorescent nucleic acid stain for quantifying double-stranded DNA.
10
Culturing Normal Human Keratinocytes for Elasticity Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Normal human keratinocyte (NHK) cultures were established from abdominal skin samples from participants who were 20 (strain 1) and 32 (strain 2) years of age according to previously published procedures (Michopoulou et al., 2020 (link)). The cultured cells were grown in a keratinocyte growth medium supplemented with 0.15 mM CaCl2 (KBM‐2 BulletKit, Lonza Biosciences, Basel, Switzerland). To initiate experiments, 2 × 105 keratinocytes were cultured in CytoSoft® 6‐well plates, elastic moduli 2 and 32 kPa (Sigma Aldrich), for 48 h in keratinocyte basal medium (KBM)‐2. Wells were rinsed with sterile PBS and processed for RNA extraction. Cells were photographed using a Zeiss Axiovert 40 microscope equipped with a circular differential interference contrast and coupled to a Coolsnap Fx camera (Roper Scientific, Evry, France).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!