User:Rebeca Rodriguez/Notebook/581/2014/10/01

Biomaterials Design Lab

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Objective Two groups will be working on determining fluorophore concentration. (Alicia/Madeleine and Becky/Eleni/Melvin) The other two groups will be working on diffusion. (Jake/James/Mary and Andrew/Michael/Tami). NOTE: The groups working on fluorophores will need to space to the groups working on diffusion as their research is very time sensitive. Description Fluorescence Make stock concentrations (both groups can use the same solutions) 0.10uM 0.50uM 1.0uM 1.5uM 2.0uM Take UV-Vis and Fluorescence spectra of these samples. Make a calibration curve based on UV-Vis. Compare your data to some published values Make a calibration curve based on the fluorescence. In order to do this, you'll need to measure the area under the fluorescence curve, not just the fluorescence peak height. Diffusion One group will use a PVA film The other group will use a PVA/clay film Set the film between the two chambers Add liquid to each chamber One chamber gets water The other chamber gets 20ppm Malachite green Withdraw 200uL from each chamber at the 5 minute mark and place in their own eppy tube NOTE: I want the groups to coordinate so that their 5 minutes don't overlap with one another. NOTE: You'll want to use a small volume cuvette NOTE: You'll also want a 0minute measurement for each. NOTE: You'll likely have to dilute the sample from your chamber with the 20ppm. So, when you return the sample, only return the non-diluted sample. Measure the UV-Vis spectrum for both samples. Return each to the chamber it originally came from Take measurements at the following times 15 minutes 30 minutes 1 hour 2 hours right before you leave lab It would be great if someone could stop by on Thursday to take a measurement then. You'll be taking measurements on Friday too. During each of this, it would be best to start working up the data right away. Data Fluorescents 0.0120g of R6G in 10mL needed to make a 2500uM stock solution. This solution was used to make 2.0uM. The solution was diluted further (1:5), to create a 500uM stock solution in order to make the dilutions for 0.1, 0.5, 1.0, and 1.5. Stock Concentrations (uM) Amount of Stock Solution Needed (mL) 0.1 0.001 0.5 0.005 1 0.01 1.5 0.015 2 0.004 Fluorescent Parameters Value Exitation Wavelength 500 nm Range 515nm -700nm Slit Width 10nm Scan Speed 200 nm/min Notes This area is for any observations or conclusions that you would like to note. Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.