Wound scratch and transwell assays were performed to evaluate cell migration and invasion, respectively. For the wound scratch assay, cells were seeded in a 6-well culture plate and allowed to grow to confluence. The cell monolayer was scratched with a sterile pipette tip and washed with PBS to remove the debris. The cells were then incubated in serum-free medium for 24 h, and wound closure was observed and photographed under an inverted microscope (Olympus, Tokyo, Japan). For the transwell assay, cells were seeded in the upper chamber of a transwell insert (Corning Inc., Corning, NY, USA) with or without Matrigel (BD Biosciences, San Jose, CA, USA) coating. The lower chamber was filled with medium containing 10% FBS as a chemoattractant. After 24 h of incubation, the cells that migrated or invaded through the membrane were fixed with 4% paraformaldehyde and stained with crystal violet. The cells were then observed and photographed under an inverted microscope (Olympus, Tokyo, Japan).
Inverted microscope
Manufactured by Olympus
Sourced in Japan, United States, Germany, China, United Kingdom, India, Italy
The Olympus Inverted Microscope is a versatile optical instrument designed for the observation and analysis of samples. It features an inverted configuration, with the objective lenses positioned below the specimen stage, allowing for the examination of cell cultures, microplates, and other samples that require an upright orientation. The inverted design provides a more convenient and accessible workspace for the user.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (235 mentions)
Transwell chamber, by Corning (121 mentions)
Matrigel, by Corning (103 mentions)
Crystal violet, by Merck Group (61 mentions)
Transwell chamber, by BD (30 mentions)
Transwell insert, by Corning (26 mentions)
Matrigel, by Merck Group (25 mentions)
Crystal violet, by Beyotime (25 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (25 mentions)
Paraformaldehyde (pfa), by Merck Group (22 mentions)
Matrigel matrix, by BD (19 mentions)
24 well transwell chamber, by Corning (19 mentions)
Crystal violet, by Solarbio (17 mentions)
Transwell, by Corning (16 mentions)
Upper chamber, by Corning (15 mentions)
Transwell chamber, by Merck Group (15 mentions)
Oil red o, by Merck Group (13 mentions)
Transwell plate, by Corning (13 mentions)
Crystal violet, by Sangon (12 mentions)
Paraformaldehyde (pfa), by Solarbio (12 mentions)
2 417 protocols using inverted microscope
1
Evaluating Cell Migration and Invasion
2
Wound Healing and Cell Migration Assays for Papillary Thyroid Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After transfection for 36 h, TPC-1 and IHH-4 cells were starved for 6 h and then a scratch was created in the bottom of each well using a 200 μL-pipette tip. The remaining cells were cultured in serum-free medium for another 24 h. The wounds were captured at 0 and 24 h under an inverted microscope (Olympus, Tokyo, Japan), and analyzed on Image J v1.46 software (National Institute of Health, Bethesda, MD, USA). Eventually, the wound healing rate was calculated using 100% × (Width 0h -Width 24h )/Width 0h .
The starved cells (5 × 10 5 cells) were re-suspended in The role of circRPS28 in PTC 200 μL serum-free medium, followed by the cell inoculation into the upper place of Transwell chamber (Corning, Carlsbad, CA, USA) for cell migration assay. For invasion assay, the chambers were pre-coated with 50 μL Matrigel (#354480; BD Biosciences) overnight at 4°C. 500 μL medium containing 20% FBS (R&D systems) was filled in the bottom place in 24-well plate. These chambers and plates were together incubated at 37°C for 48 h. Transferred cells were fixed with 4% paraformaldehyde (Beyotime) for 30 min and stained with 0.5% crystal violet (Beyotime) for 30 min. The migration number and invasion number were counted under an inverted microscope (Olympus) at 100×, and five randomly selected fields were used.
The starved cells (5 × 10 5 cells) were re-suspended in The role of circRPS28 in PTC 200 μL serum-free medium, followed by the cell inoculation into the upper place of Transwell chamber (Corning, Carlsbad, CA, USA) for cell migration assay. For invasion assay, the chambers were pre-coated with 50 μL Matrigel (#354480; BD Biosciences) overnight at 4°C. 500 μL medium containing 20% FBS (R&D systems) was filled in the bottom place in 24-well plate. These chambers and plates were together incubated at 37°C for 48 h. Transferred cells were fixed with 4% paraformaldehyde (Beyotime) for 30 min and stained with 0.5% crystal violet (Beyotime) for 30 min. The migration number and invasion number were counted under an inverted microscope (Olympus) at 100×, and five randomly selected fields were used.
3
Wound Healing and Cell Migration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For wound healing, in short, the transfected cells were planted in 6-well plates and cultivated to the density of 80%. A pipette tip (100 µL) was used to create scratches and the cells were incubated with serum-free medium for 24 h. Finally, the wound healing photographs were captured using an inverted microscope (Olympus, Tokyo, Japan) and the movement distance of cells was calculated using Image J. For the transwell assay, the transfected cells were planted in the upper transwell chamber supplemented with the serum-free medium, and the serum medium was added into the lower transwell chamber. After incubation at 37 °C for 24 h, these cells were immobilized and stained. Subsequently, an inverted microscope (Olympus, Japan) was utilized to take images and count the cell numbers.
4
Wound Healing and Transwell Migration Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The wound-healing migration assays were done in conjunction with siRNA transfections in control or 4-OHT-treated HKe3 ER:HRAS V12 cells. Seventy-two h after siRNA transfections, confluent monolayers of cells were wounded with a p20 pipette tip (time 0). Phase-contrast images were taken using an Olympus inverted microscope at time 0 h or 24 h after the scratch wound. For the Transwell migration assay, Transwell membranes (8-μm pore size, 6.5-mm diameter; Corning Costar, 3422) were used. The bottom chambers of the Transwell were filled with migration-inducing medium (with 50% FBS). The top chambers were seeded with 1.5 × 105 live serum-starved control or ATG5 shRNA HCT116 cells per well. After 48 h, the filters were fixed with 4% paraformaldehyde for 10 min at room temperature; subsequently, the cells on the upper side of the membrane were scraped with a cotton swab. Similar inserts coated with Matrigel (Corning, 354480) were used to determine invasive potential in invasion assays. Filters were stained with crystal violet for light microscopy. Images were taken using an Olympus inverted microscope and migratory cells were evaluated by ImageJ 1.42q software (National Institutes of Health, United States).
5
Cell Migration and Invasion Assay
6
Wound Healing and Invasion Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For wound healing assays, HCC‐LM3, MHCC‐97H, SMMC‐7721, or Bel‐7402 cells were seeded in a six‐well plate (5 × 105 cells/well). A scratch wound was then produced by scratching the surface of confluent cell layers with a 200‐μL pipette tip. At 0 and 24 h post scratching, the distance of the scratches was tested with an inverted microscope (Olympus).
For transwell invasion assays, the upper chambers (8.0 μm, Corning Costar Corp, Corning, NY) were first pre‐coated with 100 μL of 1 mg/mL of Matrigel (BD biosciences) for 2 h at 37°C. Next, after starvation in serum‐free medium for 24 h, HCC‐LM3, MHCC‐97H, SMMC‐7721 or Bel‐7402 (5 × 104) were seeded to the upper chambers. The lower chamber contained culture medium with 10% FBS (500 μL). After cells were incubated for 24 h, the invaded cells were stained with 0.5% crystal violet and visualized using an inverted microscope (Olympus). The cell invasion index was calculated by normalizing the number of invasive cells to the number of seeded cells in the upper chamber.
For transwell invasion assays, the upper chambers (8.0 μm, Corning Costar Corp, Corning, NY) were first pre‐coated with 100 μL of 1 mg/mL of Matrigel (BD biosciences) for 2 h at 37°C. Next, after starvation in serum‐free medium for 24 h, HCC‐LM3, MHCC‐97H, SMMC‐7721 or Bel‐7402 (5 × 104) were seeded to the upper chambers. The lower chamber contained culture medium with 10% FBS (500 μL). After cells were incubated for 24 h, the invaded cells were stained with 0.5% crystal violet and visualized using an inverted microscope (Olympus). The cell invasion index was calculated by normalizing the number of invasive cells to the number of seeded cells in the upper chamber.
7
Cell Proliferation and Migration Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The CCK-8 assay was performed to assess the proliferative capacity in different groups. A total of 3000 cells were seeded in each well of the 96-well plates. Then, 10 μL CCK-8 reagent (Beyotime, China) was added into each well at 24 h, 48 h, and 72 h post-transfection. After incubation at 37 °C for 1 h, the absorbance at 450 nm of each well was detected. The wound healing assay and transwell assay were performed to assess the migration capacity in different groups. For the wound healing assay, transfected cells were added to 24-well plates. Once the confluence reached 100%, a 200 μL pipette tip was used to scrape on the bottom of the culture plates. After removing the debris, the cells were incubated in serum-free medium for 24 h, then images were taken under an inverted microscope (Olympus, Tokyo, Japan). For the transwell assay, a 200 μL serum-free medium containing 5 × 104 transfected cells was added into the upper chamber (Corning, New York, NY, USA), while a 600 μL medium that contained 10% FBS was introduced into the bottom chamber. After 24 h, the upper side of the chamber was thoroughly wiped off with cotton swabs, and the migrated cells were fixed with methanol and stained with 0.1% crystal violet. More than three fields were captured randomly under an inverted microscope (Olympus, Japan).
8
Morphological Assessment of Cultured Follicles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The morphology of cultured follicles was assessed
under an inverted microscope (Olympus, Japan)
every 48 hours during culture period. The follicles
with dark appearance were defined as degenerated
ones. The follicle diameter (n=10/each group) was
determined by precalibrated ocular micrometer
(Olympus, Japan) using an inverted microscope
(magnification: ×100) during culture period.
under an inverted microscope (Olympus, Japan)
every 48 hours during culture period. The follicles
with dark appearance were defined as degenerated
ones. The follicle diameter (n=10/each group) was
determined by precalibrated ocular micrometer
(Olympus, Japan) using an inverted microscope
(magnification: ×100) during culture period.
9
Analyzing Cell Migration Abilities
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell migration ability was assessed by Transwell migration and wound healing assays. For the Transwell migration assay, BM-MSCs treated with either 10 μM CANA, 10 μM DAPA, or 0.1% (v/v) DMSO (as a negative control) for 48 h were suspended in serum-free medium at a concentration of 5 × 104 cells/mL. A 200-μL cell suspension (1 × 104 cells) was pipetted into the upper chamber. DMEM with 10% foetal bovine serum was added to the lower chamber as a chemoattractant. BM-MSCs were incubated for 24 h at 37 °C, and the cells on the upper side of the membrane were then removed with a cotton swab. The cells that migrated to the lower surface were fixed with 4% paraformaldehyde, stained with 4′,6-diamidino-2-phenylindole (DAPI) or 0.1% crystal violet, and observed using an inverted fluorescence microscope (Leica, Germany) or inverted microscope (Olympus, Japan). For the wound healing assay, BM-MSCs were cultured in 12-well plates, treated as indicated for 48 h, and allowed to grow to confluence. Linear wounds were made with a sterile 200-μL pipette tip. The cells were washed to remove debris using PBS and incubated in serum-free DMEM at 37 °C for 72 h. Serial images of cell migration at 0, 24, 48 and 72 h after scratching were obtained using an inverted microscope (Olympus, Japan). The percentage of closure of the wounded areas at 72 h was calculated as the 72-h migration ratio.
10
Wound Healing Assay in Hep3b Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Hep3b cells were inoculated in a 6-well plate (4x104 cells/well). When the confluence was up to 70–80%, a wound on the cell surface was formed by a pipette tip. An inverted microscope (Olympus Corporation) was adopted to observe the cells during the experiments and the images were labeled as 0 h. Subsequently, the medium was taken place of serum-free medium, in which a more detailed cell culture was performed for 24 h. An inverted microscope (Olympus Corporation) was utilized to photograph the cell migration.
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!