ONS-76 cells (2.5 × 10 4 ) were seeded in 6-well plates and grown until they formed a confluent monolayer. Following treatment with ribavirin or H 2 O, a scratch wound was made in the center of the well by scraping it with a P200 pipette tip. Wells were then washed with phosphatebuffered saline (Invitrogen) and incubated to allow cells to migrate into the space cleared by the scratch. Cell migration was assessed by capturing images from identical locations along each wound using a Zeiss Observer Z.1 AX10 microscope at 0 and 12 hours after the scratch.
Observer z 1 ax10 microscope
Manufactured by Zeiss
Sourced in United States
The Zeiss Observer Z.1 AX10 microscope is a high-performance laboratory equipment designed for optical microscopy. It features a modular design and supports a range of imaging techniques, including brightfield, darkfield, and phase contrast. The microscope is equipped with a high-resolution camera and advanced optics to provide clear and detailed images for scientific research and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Biocoat matrigel invasion chamber, by Corning (1 mentions)
Matrigel, by Corning (1 mentions)
Gelatin, by Merck Group (1 mentions)
Laminin, by Merck Group (1 mentions)
Anti eif4e fitc, by BD (1 mentions)
Fluoro gel 2 with dapi, by Electron Microscopy Sciences (1 mentions)
Matrigel coated insert, by BD (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
6 protocols using observer z 1 ax10 microscope
1
Scratch Wound Assay for Cell Migration
2
Cell Invasion Assay with Ribavirin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Invasion assays were performed using Corning BioCoat Matrigel Invasion Chambers. ONS-76 cells (1.0 × 10 5 ) were pretreated with ribavirin or H 2 O and plated in the top chamber in serum-free medium. Cells were allowed to invade into the lower chamber (containing culture medium with 10% fetal bovine serum) for 24 hours. The bottom of each insert was then fixed and stained with crystal violet. The numbers of invading cells were assessed using a Zeiss Observer Z.1 AX10 microscope.
3
Adhesion Assay for Ribavirin-Treated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Migration and Adhesion assays were performed as described previously [52 (link)]. For the adhesion assay, ribavirin-pretreated (48 hrs) or control cells were seeded into 24-well plates coated with gelatin (2 mg/ml, Sigma-Aldrich), Matrigel™ (10 mg/ml, Corning, NY, USA), or laminin (10 μg/ml, Sigma-Aldrich) and incubated for 1hr at 37°C. Cells were then carefully washed 3 times with PBS, fixed, and counted. The number of remaining adherent cells was counted in at least 4 different wells for each condition using a Zeiss Observer Z.1 AX10 microscope (Zeiss, Thornwood, NY, USA) and ImageJ software (Windows 1.47, Research Services Branch, NIH, Bethesda, MD, USA).
4
Quantifying eIF4E Expression in Ribavirin-Treated Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BT12 and BT16 cells (10-50×103/sample) were plated on microscope slides and treated with ribavirin (50 μM) or control for 72 hrs. Slides were then dried at 37°C, fixed and permeabilized with 100% methanol at −20°C for 20min, washed with PBS, and blocked with 10% fetal bovine serum/0.2% Tween-20/PBS at 37°C for 1hr. Slides were then labeled with 1:50 anti-eIF4E-FITC (BD Transduction) in 10% FBS/0.2% Tween-20/ PBS at 37°C for 2 hrs. Slides were then mounted with Fluoro-Gel II with DAPI (Electron Microscopy Sciences, Hatfield, PA, USA). DAPI and eIF4E images were acquired with a Zeiss Observer Z.1 AX10 microscope (Zeiss).
5
Intracranial Implantation of AT/RT Cells in Athymic Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice (NU/NU athymic) were purchased from Charles River (Cambridge, MA, USA) and intracranially implanted with 5×105 BT12 AT/RT cells as previously described [53 (link), 54 (link)]. Animals were given intraperitoneal injections (200μL) of pharmaceutical-grade anesthesia, analgesia, and study agents (water vehicle or ribavirin, 100 mg/kg/day). At day 20 after implantation, mouse brains from vehicle- and ribavirin-treated groups were collected, fixed in formalin, paraffin-embedded, sectioned, and H&E stained. Images were taken using a Zeiss Observer Z.1 AX10 microscope (Zeiss), and quantification of tumor sizes was performed using ImageJ software. All procedures were performed in accordance with the guidelines set forth by the Johns Hopkins University Animal Care and Use Committee.
6
Matrigel Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The invasion assay was performed using 24-well plates with Matrigel-coated inserts (8μm pores, BD Biosciences). Ribavirin-pretreated and control cells (1×105) were placed in the top chamber containing medium without serum. The lower chamber contained medium with 10% fetal bovine serum. After invasion, non-invasive cells were removed from the top of the insert using cotton swabs. The underside of each membrane was fixed and stained with DAPI (Invitrogen). The number of cells was counted for each condition using a Zeiss Observer Z.1 AX10 microscope (Zeiss). The data are representative of three independent experiments.
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!