For tumor spheroid formation, a total of 5000 single cell suspensions were plated in 100 μL of complete media (DMEM supplemented with 20% FBS, 1x MEM non-essential amino acids, and 1x Anti-Anti) into each well of a 96-well ultra-low attachment plates (Corning 3603). After 3 days in culture, tumor spheroid formation was confirmed visually using the EVOS XL Core Cell Imaging System (Thermo Fisher Scientific). On day 0, 100 μL of media containing 2x concentration of drug was added to the wells diluting the compound to the indicated concentration. Every 3 or 4 days, 100 μL of media was removed and replaced with 100 μL of media with 2x concentration of drug. The spheroids were treated for 21 days. The spheroids where then dissociated with Cell Titer Glo 3D (Promega # G9681) following manufacturer’s instructions. The plates were then read on an Enspire Mulitmode Plate Reader (Perkin-Elmer).
Enspire mulitmode plate reader
Manufactured by PerkinElmer
Sourced in United States
The Enspire Multimode Plate Reader is a versatile laboratory instrument designed for a wide range of detection modes, including absorbance, fluorescence, luminescence, and time-resolved fluorescence. The device supports a variety of microplate formats and can be used for various applications in fields such as drug discovery, cell-based assays, and molecular biology.
Automatically generated - may contain errors
4 protocols using enspire mulitmode plate reader
1
Tumor Spheroid Formation and Drug Screening
2
Quantitative Lipid Droplet Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lipid droplet quantification by Nile Red staining was performed after incubation of cells with serum free medium for 5 days. Nile Red (15 μg/μL) (Santa Cruz Biotechnology, Dallas, United States) was added to the cell medium for 20 min at 37°C. Nile Red fluorescence was analyzed at 580 nm using an EnSpire Mulitmode plate reader (PerkinElmer, Waltham, United States). For normalization cell vitality was assessed using the alamarBlue assay (ThermoFisher, Waltham, United States) according to the manufacturer’s instructions.
3
Tumor Spheroid Formation and Drug Screening
4
In vitro Assembly of RNA Biosensors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vitro assembly reactions were
prepared on ice
with 25 picomoles of each RNA and 250 picomoles of DFHBI-1T dye (Lucerna
Technologies) in buffer (40 mM HEPES pH 7.5, 100 mM KCl, 1 mM MgCl2) at a final volume of 50 μL. For thermal renaturation,
samples were transferred into a preheated aluminum insert within a
dry heat block set to 90 °C. Following addition of samples, the
aluminum insert was immediately removed from the block heater and
placed on the lab bench to cool to 37 °C before measurement.
Assembly at 37 °C was performed by placing samples in a dry heat
block at 37 °C for 15 min before measurement. Samples were then
transferred into a clear, flat-bottom 96-well plate and measured for
fluorescence (λex = 472 nm, λem =
507 nm) on an EnSpire Mulitmode plate reader (PerkinElmer) at room
temperature. Measurements from No RNA samples, which
contained buffer and dye, were averaged to establish background signal.
Background signal was subtracted from all measurements and individual
readings were normalized for fluorescence relative to 3WJdB. Normalized values were used to calculate mean and standard deviation.
prepared on ice
with 25 picomoles of each RNA and 250 picomoles of DFHBI-1T dye (Lucerna
Technologies) in buffer (40 mM HEPES pH 7.5, 100 mM KCl, 1 mM MgCl2) at a final volume of 50 μL. For thermal renaturation,
samples were transferred into a preheated aluminum insert within a
dry heat block set to 90 °C. Following addition of samples, the
aluminum insert was immediately removed from the block heater and
placed on the lab bench to cool to 37 °C before measurement.
Assembly at 37 °C was performed by placing samples in a dry heat
block at 37 °C for 15 min before measurement. Samples were then
transferred into a clear, flat-bottom 96-well plate and measured for
fluorescence (λex = 472 nm, λem =
507 nm) on an EnSpire Mulitmode plate reader (PerkinElmer) at room
temperature. Measurements from No RNA samples, which
contained buffer and dye, were averaged to establish background signal.
Background signal was subtracted from all measurements and individual
readings were normalized for fluorescence relative to 3WJdB. Normalized values were used to calculate mean and standard deviation.
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!