Questions to ask the fluorometer Engineer on Friday, Aug 3
See if our plate reader has the ability to measure what you want to measure.
- What are the classes of measurement? Can it do time-resolved fluorometry + Absorbance?
- Delay time-resolve - 24 hour experiment; evaporation of solution over long-time courses?!
- How do we subtract background values from data and convert from absorbance to OD600?
Talk about using the plate reader and creating protocols.
- Excitation filters and Emission filters for GFP, acGFP and DsRed-Express?
- Any manufacturer protocols for plate reader usage?
- Well plate rows and columns? How is data arranged sequentially with the wells?
- What kind of plates you need to order and use for our experiments?
- Lamp energy and Noise signal ratios??
Talk about processing data
- Export format?
- How to export data from fluorometer?
- Built-in data analysis tools?
- Technical Support.
Experiment 1: Fluorescence vs extracellular [GFP] molecules
Known concentrations of GFP are diluted into a range of dilutions and are then mixed with cell lysate of a particular chassis.
Experiment 2: Fluorescence vs Cultures of varying unknown intracellular [GFP]
Measuring the fluorescence of cultures of cells at set time intervals would measure a set of unknown intracellular [GFP].
Experiment 3: Fluorescence vs Cultures of varying unknown extracellular [GFP]
Lysing samples of Experiment 2 will allow the relation of an unknwon intracellular [GFP] to that of extracellular [GFP].