For the examination of the morphology change during EMT process, treated U2OS and HOS cells were collected and seeded into 24-well plates at a concentration of 2 × 106 cells per well. The cell morphology was monitored by an inverted microscope (Axiovert 10 ZEISS).
Axiovert 10
Manufactured by Zeiss
Sourced in Germany
The Axiovert 10 is an inverted microscope designed for routine observation and documentation. It features a high-quality optical system and a stable mechanical construction. The Axiovert 10 is suitable for various applications, including cell culture monitoring, simple brightfield, and phase contrast observation.
Automatically generated - may contain errors
Lab products found in correlation
Versamax microplate reader, by Molecular Devices (2 mentions)
Rat tail collagen, by Merck Group (1 mentions)
Dabco, by Carl Roth (1 mentions)
Mowiol, by Merck Group (1 mentions)
Matrigel growth factor reduced basement membrane matrix, by Corning (1 mentions)
Matrigel, by Zeiss (1 mentions)
96 well culture plate, by BD (1 mentions)
Axoclamp 2a amplifier, by Molecular Devices (1 mentions)
Labchart 7, by ADInstruments (1 mentions)
Powerlab system, by ADInstruments (1 mentions)
Vectashield solution, by Vector Laboratories (1 mentions)
Axopatch 200b, by Molecular Devices (1 mentions)
Bcecf am, by Thermo Fisher Scientific (1 mentions)
Aec 3 amino 9 ethylcarbazole staining kit, by Merck Group (1 mentions)
Avicel rc 581, by FMC Biopolymer (1 mentions)
25 protocols using axiovert 10
1
Monitoring EMT Cell Morphology
2
Immunofluorescent Staining of Virus-Infected CRFK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CRFK cells were cultured on sterile cover slips in 24 well plates pretreated with rat tail collagen (Sigma). Cells were mock or virus-infected and treated with 2 μg ml−1 tunicamycin 4 h p.i. At 15 h p.i. the cells were fixed with PBS containing 4% paraformaldehyde for 20 min at 4 °C, followed by three PBS washes. Cells were permeabilized with Triton X-100 (1% in PBS) for 5 min at room temperature. The cells were rinsed twice with PBS and residual binding sites were blocked with 1× Roti®-ImmunoBlock (Roth) for 10 min at room temperature. The cells were then incubated with primary antibodies (1:3 dilution) or sera (1:1500 dilution) in PBS for 1 h at 37 °C, followed by three PBS washes. Subsequently the cells were incubated with secondary antibodies conjugated to cyanogen-3 (1:500 dilution, Dianova) in PBS for an additional hour at 37 °C. The DNA was counterstained with 66 μg ml−1 4′,6-diamidino-2-phenylindole (DAPI) for 5 min at room temperature. After three washes the cover slips were mounted with Mowiol (Sigma) containing the anti-fading reagent DABCO (1,4-diazabicyclo(2.2.2)octane, Roth). The immunofluorescent staining was visualized by fluorescence microscopy (Axiovert 10, Zeiss).
3
Cell Migration Assay with DZ-50
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wounding was inflicted using a sterile pipette tip in confluent cell monolayers in 6-well plates. After incubation for 12–48 hrs in the presence of DZ-50 (5 µM), wounded areas were examined by light microscopy (Axiovert 10, Zeiss). Cells migrating to the wounded areas were counted under a microscope. Migration potential was determined as the average number of cells in three random high-power (400×) fields/well. Numerical data are obtained from three independent experiments performed in triplicate and is expressed relative to untreated controls.
4
Angiogenesis Assay with EAhy926 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EAhy926 cells were seeded into a 6-well plate and cultured in full media until they reached confluency. The cells were serum starved for one hour, washed with PBS, and harvested. Matrigel (Corning Matrigel Growth Factor Reduced Basement Membrane Matrix; Corning, NY14831, USA) was diluted 1:1 with serum-free media and prewarmed at 37 °C in a 96 well plate. After seeding with approximately 85000 cells in serum free media on the Matrigel, the cells were incubated at 37 °C for 24 h in medium with DMSO ± PSP-2 at a final concentration of 5 µM. Two to three images per well were captured at 10x magnification, using a phase contrast inverted microscope (Axiovert-10, Carl Zeiss AG, Feldbach, Switzerland), equipped with a digital camera. Images were analyzed using the Angiogenesis Analyzer, a plugin developed for the ImageJ (Version 1.47t; NIH, Bethesda, MA, USA).[ref]
5
Antiviral Efficacy Evaluation by SRB Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antiviral activity was assessed by the SRB method using cytopathic effect (CPE) reduction as reported previously (Song et al., 2014 (link)). Briefly, one day prior to infection, Hela cells (2×104 cells/well) were seeded onto a 96-well culture plate (BD biosciences, San Jose, CA, USA). On the next day, medium was replaced with medium containing 30 mM of MgCl2, 1% FBS, diluted virus suspension containing a 50% cell culture infective dose (CCID50) of the virus, and an appropriate concentration of the test compounds. The culture plates were incubated at 32°C in 5% CO2 for 2 days until the appropriate CPE was achieved. After incubation in ice-cold 70% acetone for 30 min, cells were stained with 0.4% (w/v) SRB in 1% acetic acid solution. Cell morphology was observed using an Axiovert microscope (Axiovert 10; Carl Zeiss, Oberkochen, Germany) to examine the effect of the compounds on HRV-induced CPE. Bound SRB was then solubilized with 10 mM unbuffered Tris-based solution, and absorbance was read at 562 nm using a VERSAmax microplate reader (Molecular Devices, Palo Alto, CA, USA) with reference absorbance measured at 620 nm. The percentage of cell viability was calculated for comparison based on the measured absorbance. In addition, the cell morphology was observed under a microscope at 32×10 magnification (St Ernst-Leitz, Wetzlar, Germany), and images were recorded.
6
Measuring Action Potential of Beating Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The action potential of the beating SVF cells was recorded on day 28, as described previously45 (link). Briefly, the action potential was recorded using patch pipettes at 37℃ with an external control solution containing (in mmol/L) NaCl 140, KCl 5.4, CaCl2 1.8, MaCl2 2, glucose 10, and HEPES 10 (pH 7.4). Suction pipettes were generated from borosilicate capillary tubes (8250 glass; A-M Systems) and fire-polished to obtain a resistance of 5–8 MΩ when filled with a solution containing (in mmol/L) KCl 120, EGTA 5, K2ATP 5, MgCl2 2.5, and HEPES (pH 7.2). Under the Axiovert 10 (Carl Zeiss, Jena, Germany) microscope, Vm was measured using the AxoClamp 2A amplifier (Molecular Devices) in bridge mode via the disrupted patch technique, as described previously43 (link). Vm was digitised using the PowerLab system (AD Instruments, Dunedin, New Zealand) and recorded at a sampling frequency of 2 kHz/16-bit in LabChart 7.0 (AD Instruments) using a 0.5–40 Hz band-pass filter. All recording equipment was housed on an air table within a grounded Faraday cage to minimise background noise. The APD50 and APD90 of repolarisation were calculated from a series of 10 potentials.
7
Essential Oils Inhibit Influenza Virus CPE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effect of essential oils on influenza virus-induced CPE was observed. Briefly, MDCK cells were seeded onto a 96-well culture plate at a concentration of 2 × 104 cells per well. The next day, the culture medium was removed and the cells were washed with PBS. Diluted virus suspension (0.09 mL) and 0.01 mL of medium supplemented with trypsin-EDTA containing essential oils at 100 μg/mL were added to each well. After incubation at 37°C in 5% CO2 for 2 days, the morphology of cells was observed under the microscope at 32 × 10 magnifications (AXIOVERT10, ZEISS, Germany), and images were recorded.
8
Conditioned media preparation and effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To prepare conditioned media (CM), LC212 and LC31 cells [22 (link)] were cultured in standard medium at a density of 200.000 cells/fl 25. After 24 h the cells were washed and cultured with RPMI deprived of FBS for 48 h. To remove cells and cell debris, the collected media were centrifuged for 10 min at 14,000 rpm and 4°C, and supernatants were used as conditioned media study. A549 cells were cultured at a density of 75.000 cells/well in six wells. After 48h of culture in the standard medium, A549 cells were cultured for 96 hours with LC212 and LC31-CM, and with standard medium alone. Cell morphology was captured using an inverted microscope (Axiovert 10 ZEISS).
9
Immunofluorescence Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with 4% paraformaldehyde in PBS for 10 min at room temperature, then permeabilized in 0.05% Triton X-100 in PBS and incubated with blocking solution (2% glycine, 2% BSA, 5% FBS, 50 mM NH4Cl in PBS pH 7.4) for 1 h at room temperature (RT). After incubation with appropriate primary (4 °C) and secondary antibodies (RT), coverslips were mounted with Vectashield solution (Vector, Burlingame, CA, USA). Images were acquired with a Zeiss Axiovert 10 epifluorescence microscope furnished with 40× and 100× Zeiss Apoplan oil immersion objectives and equipped with a Retiga-SRV camera operated with the standard QC capture software (Q-Imaging). Quantification was performed with ImageJ software (NIH, USA) using the corrected total cell fluorescence (CTCF) method, CTCF = integrated density – (area of selected cell × mean fluorescence of background readings), where CTCF is the corrected total cell fluorescence. Both the endogenous and recombinant protein levels were quantified, and the data were normalized to arbitrary units (A.U.).
10
Epi-Fluorescence Microscopy Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This system is based on an epi-fluorescence inverted microscope (Axiovert 10, Zeiss, Jena Germany). The excitation light from a Xenon lamp passed through bandpass filters mounted on a computer-controlled rotating wheel that allows alternate stimulation at 340 ± 10 and 380 ± 10 nm with a frequency of 3.3 Hz. The excitation light was deflected by a dichroic mirror (FT 425, Zeiss, Jena, Germany) through an oil-immersion objective (Plan Neofluar 100 × 1.30, ph. 3, Zeiss, Jena, Germany). Fluorescence emission from individual cells was spatially limited by a diaphragm adjusted to the cell size (10–15 μm).
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!