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Unformatted text preview: Bio 1AL Lab Lecture Outline # 1 January 25 th , 2010 I. Biological Themes A. High levels of structural organization. atoms -> molecules -> macromolecules -> organelles-> cells-> tissues-> organs -> organ systems -> organism B. Making order of diversity – use of taxonomy. D > K > P > C > O > F > G > S C. Prokaryotes versus eukaryotes – presence of membrane bound organelles ("nucleus") D. Three Domains: Eubacteria, Archae, Eukarya II. Micropipetter A. Why – measure small volumes precisely B. Use – tips & points of resistance i) P20 – (1-20 μ L), P200 – (20-200 μ L), P1000 – (200-1000 μ L) C. Do's & Don'ts D. Sterile technique E. Exercise – weigh various volumes w/ a balance III. Microscope A. Why? – increase magnification/resolution B. Parts – imaging and illumination. C. Optical Sectioning D. Critical Illumination – condenser position, amt. of light E. Exercises 1. field of view diameter – parachute cloth 2. "ruler" calibration – parachute cloth 3. letter e 4. depth of field - solid glass rod, hollow tube. 1) Stage down and Condenser down -> 2) Rotate intensity to low, power on and mount slide – position as necessary (look from the side) -> 3) Focus (coarse and fine) with right eye only -> 4) Use left eye only and adjust diopter as necessary -> 5) Adjust interpupillary distance (one field of view) -> THIS ONLY NEEDS TO BE DONE ONCE PER SLIDE. Turning on, Light Control Diopter adjustment (both eyes in focus) Set Interpupillary Distance 6) Close field diaphragm, adjust condenser position until edge of field diameter is in best focus -> 7) Center field diaphragm if necessary -> 8) Adjust field diaphragm size -open field diaphragm until it is slightly larger than field of view-> 9) Increase magnification ->10) Adjust field diaphragm size and position condenser (repeat 6 & 8) -> 11) Fine focus as necessary -> 12) Adjust aperture diaphragm as necessary 12) Increase magnification- repeat 10-12. Critical Illumination – Size of field diaphragm, position of condenser, size of aperture diaphragm. 1) Condenser up, Stage Down, Mount slide (see image in path of light) 2) Focus, adjust diopter, set interpupillary distance 3) Center (only once) and size field diaphragm and set position of condenser. Use aperture diaphragm as necessary. 4) Fine focus only at higher magnifications. VI. Cells A. Optical sectioning. B. Prokaryotes vs. Eukaryotes – presence of membrane bound organelles "nucleus" C. Far more similarities than differences! D. Size limits- Increasing cell size. i) volume increases faster than surface area. ii) rates of diffusion – bulk mixing of fluids iii) dilution of reactants/products – compartmentalization can help (Eukaryotes) E. Prokaryotes vs. Eukaryotes i) Prokaryote structure Fig. 6.6 [7 th & 8 th ed.] ii) Gram staining iii) Eukaryote structure (plant cell) Fig. 6.9 [7 th & 8 th ed.] F. Drawing guidelines – making a drawing VI. Sea water analysis – isolation of bioluminescent bacteria....
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- Spring '08
- DNA, Aperture, field diaphragm, aperture diaphragm