SHG44 and U251 cells in their log growth phase were passaged and transferred into the wells of a six-well plate at a density of 5×105 cells/mL. The cells were then cultured for 48 hours in a 37°C incubator with 5% CO2. Straight scratches were created on the bottom of the wells with a 10 µL pipette tip. PBS solution was used to wash off suspending cells, and the remaining cells were cultured in serum-free medium for another 48 hours. A MOTIC inverted microscope was used for observation and photography, and IPP software was used to analyze the width of the scratches.
Inverted microscope
Manufactured by Motic
Sourced in Canada, China, Germany, Hong Kong, United States
The Motic inverted microscope is a type of microscope that allows for the observation of specimens from below. This design enables the examination of living cells and tissues in their natural states, as the illumination and objectives are positioned beneath the stage. The inverted microscope is a versatile tool used in various fields, including cell biology, tissue culture, and material science.
Automatically generated - may contain errors
Lab products found in correlation
Transwell insert, by Corning (3 mentions)
Matrigel, by BD (3 mentions)
Mitomycin c, by Merck Group (2 mentions)
Paraformaldehyde (pfa), by Sinopharm (2 mentions)
Images plus 2, by Motic (2 mentions)
Transwell chamber, by Corning (2 mentions)
Coralite594 conjugated goat anti rabbit igg h l, by Proteintech (1 mentions)
Hrp conjugated affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
Hrp conjugated affinipure goat anti mouse igg h l, by Proteintech (1 mentions)
Matrigel, by Corning (1 mentions)
Matrigel basement membrane matrix, by BD (1 mentions)
Moticam 2500 camera, by Motic (1 mentions)
Trypan blue solution, by Beyotime (1 mentions)
Facscalibur flow cytometer, by BD (1 mentions)
Costar 3595, by Corning (1 mentions)
Ae2000, by Motic (1 mentions)
Air objective, by Olympus (1 mentions)
Mowiol 4 88 mounting medium, by Merck Group (1 mentions)
Crystal violet, by Solarbio (1 mentions)
Superfrost plus, by Thermo Fisher Scientific (1 mentions)
25 protocols using inverted microscope
1
Wound Healing Assay with SHG44 and U251 Cells
2
Immunofluorescence Imaging of Cellular Targets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reagents were rabbit secondary antibody (CoraLite594-conjugated Goat Anti- Rabbit IgG (H + L), Proteintech, Rosemont, IL, USA, article number SA00013-4), rabbit secondary antibody (CoraLite594-conjugated Donkey IgG (H + L), Proteintech, Rosemont, IL, USA, article number SA00013-8), goat secondary antibody (Fluorescein (FITC)-conjugated Affinipure Donkey Anti-Goat IgG (H + L), Proteintech, Rosemont, IL, USA, article number SA00003-3), mouse secondary antibody (horseradish peroxidase (HRP)-conjugated AffiniPure Goat Anti-Mouse IgG (H + L) Proteintech, Rosemont, IL, USA, article number SA00001-1), and rabbit secondary antibody (HRP-conjugated AffiniPure Goat Anti-Rabbit IgG (H + L), Proteintech, Rosemont, IL, USA, article number SA00001-2).
Instruments included an inverted microscope (Motic, Wetzlar, Germany), fluorescence microscope, MDF-382E ultralow temperature refrigerator (Sanyo, Osaka, Japan), TONO-Pen AVIA tonometer (Medtronic SALON, USA), and HE staining kit (Bioswamp).
Instruments included an inverted microscope (Motic, Wetzlar, Germany), fluorescence microscope, MDF-382E ultralow temperature refrigerator (Sanyo, Osaka, Japan), TONO-Pen AVIA tonometer (Medtronic SALON, USA), and HE staining kit (Bioswamp).
3
In vitro Cytotoxicity of Disinfectants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All disinfectants were evaluated for in vitro cytotoxicity using PAM cells at 1:200 to 1:51,200 disinfectant dilution. Briefly, each diluted disinfectant was added to PAM cells, which were incubated for 1 h, before MM with 2% pig red blood cells was added and incubated at 37°C in a 5% CO2 incubator. The number of viable cells was determined under an inverted microscope (Motic, Fujian, China). The cutoff of cell viability was determined when live cells clung to the bottom of the tissue culture microplate for more than 80% of the 5 days culture period, equivalent to the cell control well.
4
Cell Morphology Analysis on COL1 Plates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To ensure proper comparison between experiments, both cell lines were seeded at a 0.3M cells/mL in DMEM on COL1 plates. Six hours after seeding, cells were examined though an inverted microscope (Motic, Richmond, BC, Canada) and cells showing an elongated morphology were counted in 10 different fields per well at a 400x magnification.
5
Transwell Cell Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell inserts and Matrigel were purchased from Corning Incorporated (Corning, NY, USA) and BD Biosciences (San Jose, CA, USA), respectively. We added a total of 4 × 103 cells in 200 µl cell medium into Transwell inserts that were precoated with Matrigel, and then added 800 µl medium supplemented with 30% FBS into the lower chambers. Thereafter, the cells were transfected and cultured in a cell incubator and allowed to invade for 24 hours. We removed cells on top of the membranes after rinsing. Cells (invaded through the membranes) were treated with 4% paraformaldehyde at room temperature for 20 min and stained by 0.5% crystal violet for 5 min. An inverted microscope (Motic Instruments, Richmond, BC, Canada) was applied in capturing images with a magnification of ×200.
6
Wound Healing Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells were seeded in 6-well plates (4 × 105 cells/well). When the cell confluence reached 80% after 24 hours, the cells were treated with mitomycin C (1 µg/ml; Sigma-Aldrich; Merck KGaA) at 37 °C for 1 h. Then, we created wounds on the surface of the cell monolayer by using a 200-µl pipette tip. Cells were cultured in a cell incubator and an inverted microscope (Motic Instruments, Richmond, BC, Canada) was used for image acquisition with magnification of ×100 at 24 h. The relative migration ratio was also calculated [Relative migration ratio = (incipient gap between two edges- migrated gap between two edges)/incipient gap between the two edges].
7
Transwell Invasion Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell was conducted to measure invasion ability at 48 h after transfection. The transwell insert was coated with Matrigel Basement Membrane Matrix (BD, USA) at 37°C for 1 h. The cells (5 × 104) in FBS-free RPMI-1640 were added to the insert, which was plated into a 24-well plate. The on-invasive cells were wiped out after being incubated at 37°C for 24 h, while the invasive cells were fixed and stained at room temperature. An inverted microscope (Motic, China) was used to take pictures.
8
PDGF-induced wound healing assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with PDGF for 48 h after siRNA transfection. Subsequently, the cells were cultured to ~90% confluence in serum-free DMEM containing 1 µg/ml mitomycin C (Sigma-Aldrich; Merck KGaA) at 37°C for 1 h before they were subjected to wound healing assay. Briefly, a 200 µl pipette tip was used to scratch the cell layers. Subsequently, the wound was washed with serum-free medium and further cultured in serum-free medium for 0, 12 and 24 h at 37°C. The cells were imaged under an inverted microscope (magnification, ×100) (Motic Incorporation, Ltd., Causeway Bay, Hong Kong).
9
Anticancer Effects of Cardoon Extracts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MDA-MB-231 cells (9,200 cells per well) were seeded in 0.3% agar on top of 0.5% pre-solidified agar, in a 12-well plate. Each treatment was performed in triplicate. After 24 h of incubation at 37 °C, cells were treated with cultivated cardoon leaves lipophilic extract (IC50 10.39 µg/mL) and cynaropicrin (IC50 6.19 µg/mL). Solvent control cells received DMSO (0.09% (v/v)). Every other day, agar layers were supplemented with the above-mentioned samples. After 14 consecutive days, cells were incubated overnight with 0.1% p-iodonitrotetrazolium violet at 37 °C. Colonies were thereafter observed using an inverted microscope (Motic, Xiamen, China) at the 40× magnification, and four fields of each well were photographed using Moticam 2500 camera (Motic, Xiamen, China). Images were processed using the Motic Images Plus 2.0 software (Motic, Xiamen, China).
10
Cell Seeding Density Optimization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The small volume required for seeding the cells was confined into delimited areas of the coated samples fixed by a Viton (fluorinated elastomers) O-ring because of the SH repellence of the medium culture, to avoid the sample floating in the Petri dish.
Cells were seeded at two different cell densities (2 × 105 cells/ mL and 2 × 106 cells/mL). Cells were incubated for 48 h under standard conditions. Optical microscopy through a Nikon inverted microscope equipped with a video camera (Moticam 1080 HDMI and USB) was used to monitor in situ the cell morphology and growth. Images were analyzed with the image processor Motic Images 3.0 software, Motic (Xiamen, China).
Cells were seeded at two different cell densities (2 × 105 cells/ mL and 2 × 106 cells/mL). Cells were incubated for 48 h under standard conditions. Optical microscopy through a Nikon inverted microscope equipped with a video camera (Moticam 1080 HDMI and USB) was used to monitor in situ the cell morphology and growth. Images were analyzed with the image processor Motic Images 3.0 software, Motic (Xiamen, China).
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!