For GI50 determinations, cells were seeded in 96 well plates. The next day, cells were treated with increasing concentrations of inhibitor or with DMSO alone. After a 96 h incubation period, cell proliferation was quantified by fluorescent detection of the reduction of resazurin to resorufin by viable cells and normalised to DMSO treated wells. GI50 values were calculated using non-linear regression analysis in GraphPad Prism software. For longer-term colony assays, cells were seeded into 12 well plates (0.5-5x104 cells/well), treated with inhibitors the next day and incubated for 10-14 d. Cells were fixed with 4 % formaldehyde/PBS for 20 minutes, then stained with 0.5 % crystal violet in 70 % ethanol. Excess dye was removed by washing with water and plates were imaged with a GelCount (Oxford Optronix). Cell proliferation was quantified by solubilisation of crystal violet dye in 10 % acetic acid and absorbance measured at 595 nm using an EMax Plus plate reader (Molecular Devices), then expressed as a percentage of vehicle-treated cells.
Emax plus plate reader
Manufactured by Molecular Devices
The EMax Plus is a microplate reader designed for high-performance absorbance detection. It is capable of measuring absorbance across a wide wavelength range, making it suitable for a variety of common laboratory assays.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using emax plus plate reader
1
Quantifying Cell Proliferation Inhibition
2
Broth Microdilution Assay for Sporothrix
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The broth microdilution technique, adapted for Sporothrix cells, was used to determine the minimum inhibitory concentration (MIC) values of the most active compounds [11 ]. Briefly, serial 2-fold dilutions of the compounds were prepared in supplemented RPMI in flat-bottom 96-well microplates to obtain a final concentration ranging from 0.002 to 1 µM (except for luliconazole, whose final concentration ranged from 1 to 0.0002 µM). Yeasts were added to microplates at a final concentration of 1 × 105 CFU/mL and incubated at 35 °C for 48 h in a 5% CO2 chamber. Fungal growth was analyzed by visual inspection using an inverted light microscope (Axiovert 100, ZEISS) and quantified by spectrophotometric readings at 492 nm (Emax Plus plate reader, Molecular Devices). Negative controls were included in experiments for the subtraction of absorbance values. Concentrations that inhibit at least 50% of fungal growth (IC50) were calculated using the following equation: I = 100 − (A × 100/C), where A was the absorbance of treated wells, and C was the absorbance of untreated wells. Results are presented as the mean of two independent experiments performed in duplicate.
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!