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Extra info for Lecture Notes for Methods in Cell Biology
0? Although Tris is a base the calculations are the same. Make sure that the base form and the conjugate acid form are correctly identified. 5 l = 20 g ADDITIONAL PRACTICE PROBLEMS Buffer Mol. Wt. 0? 76 and molecular weight of acetic acid is 60. 4? 5 and the molecular weight is 238. 8? 3 and the molecular weights of Tris and TrisHCl are 121 and 157 respectively. How many grams of MES (mol. wt. = 195) and the Na+ salt of MES (mol. wt. 0. 15? How many grams of Tricine (mol. wt. 5. 15? 51 52 CHAPTER 6--CENTRIFUGATION The centrifuge is an essential instrument in cell and molecular biology research.
FLUORESCENCE MICROSCOPY Since many fluorescent molecules emit light in the visible range it is possible to view fluorescence in conjunction with microscopy. An UV light source is used to illuminate the sample through the objective lens using a beam-splitting mirror. The fluorescence emitted from the sample (epifluorescence) passes through this same mirror, but the UV light does not. Filters before the beamsplitting mirror will control the excitation wavelength and filters before the eyepiece (or camera) will control the wavelength of the emitted light.
3H) are in channel A. 2. Determine the cpm in both channels A and B for all of the samples as well as a 3H standard and a 14C standard. (Standards should be counted under same conditions as samples. ) 3. Use the channel ratios of the standards to calculate the amount of each isotope in each channel. 416)(800) = 333 cpm 3 H in channel A = 500 - 333 = 167 Total 3H = 167 cpm Total 14C = 800 + 333 = 1133 cpm *In other words, at these channel settings there is no significant amount of 3H in channel B and all of the radioactivity can be assumed to be 14C.