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Understanding Biological Tools and Instrumentation: Key Techniques and Methods

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wyome
37 diapositives · 536 Views · Oct 14, 2014

This chapter explores essential biological tools and methodologies used in scientific research. It covers the systematic approach to problem-solving using the scientific method, detailing the steps from defining a problem through hypothesis testing to drawing conclusions. The types of experiments, including controlled, trial and error, and open-ended experiments are discussed. Additionally, it highlights various microscopy techniques, separation methods like centrifugation and chromatography, and the application of staining techniques and indicators in biological analysis.

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  1. Chapter 3 Biological tools and Instrumentation

  2. Taxonomic (Dichotomous) Key 1a. Is the organism unicellular -go to step 2 1b. Is the organism multicellular -go to step 3 2a. Is the organism prokaryotic -kingdom Monera 2b. Is the organism eukaryotic -Kingdom Protista

  3. Scientific Method • An systematic approach to problems solving: 1. Defining the problem “ By doing Background Research” 2. Formulating a hypothesis 3. Testing the hypothesis 4. Recording observations 5. State a conclusion

  4. Three types of Experiments • ****Controlled Experiments**** • Trial and Error • Open Ended

  5. Defining the problem “By doing Background Research” • Hypothesis – • Experimentation 1. Control group (no change) 2. Experimental group (make change) control = variable = *State a conclusion

  6. Defining the problem • Hypothesis –a possible explanation for events. • Experimentation (Testing the hypothesis) 1. Control group (no change) a source for comparison 2. Experimental group (contains 2 variables) *Independent Variable (Manipulated) *Dependent Variable (responding) #’s • State a conclusion

  7. Theory- explanation supported by experimental evidence. • Scientific Law- is a statement that describes some aspect of a phenomena that is always true.

  8. Microscopes • Simple Microscopes • Dissecting microscope • Compound Microscope • Phase Contrast Microscope • Electron Microscope • SEM • TEM

  9. Instrumentation A. Simple microscope - magnifying glass 1 –uses 1 lens B. Compound light microscope 1- uses two lenses a) ocular (eyepiece) 10x b) objective - low power, 10x - high power, 40x

  10. Compound Light Microscope • Magnifies 50x to 500x the original size • Used to look at the whole cell and organelles like the nucleus and chloroplast

  11. Adjustments: • Coarse adjustment – used to focus under low power • Fine adjustment – used to focus under high power ONLY • Used to “fine tune” the image

  12. Total Magnification: Magnifications: ocular = 10x low power objective = 10x high power objective = 40x Formula: ocular x objective = total magnification High power magnification is? 10 x 40 = 400x

  13. Imagery: • When placing a slide under the objective lens, the image appears: -magnified -upside down -inverted (backwards). Ex: e  F 

  14. e  F  F e Imagery:(as seen under low power)

  15. Imagery:Field of View (fov) What is the size of the “F”? F F Scanner fov = 4000 um Low fov=2000 um

  16. F Imagery:Field of View (fov) 4000 um

  17. Electron Microscopes • Advanced microscope magnifies 250,000x the original size • Allows for detailed observation of smallorganelleswithin the cell. • Like the mitochondria and lysosomes.

  18. Dissecting Microscope • Gives the viewer a 3-D image of the organism • Used in the dissection of small animals and plants • Low magnification • 1-10x

  19. Separation Techniques Centrifuge Chromatography Gel electrophoresis Dialysis Tubing Dissection and micro dissection instruments Stains Indicators System International Other Biological Tools

  20. Centrifuge:Separates of solutions based on each parts density Chromatography: Separates Solute from Solvent Electrophoresis:separates macromolecules-either nucleic acids or proteins-on the basis of size, electric charge, and other physical properties Dialysis Tubing Separation Techniques

  21. Centrifuge • Separates of solutions based on each parts density

  22. Gel Electrophoresis

  23. Electrophoresis Opposite charges at each side causing sample to move • Smaller DNA fragments move faster and further

  24. IV. Electrophoresis

  25. separates nucleic acids or proteins-on the basis of size, electric charge, and other physical properties Electrophoresis:

  26. Chromatography Rf= • Separates Solute from Solvent Distance traveled by the solute (ink) Distance traveled by the solvent (liquid)

  27. Thin-Layer Chromatography (TLC)

  28. Chromatography

  29. Retention Factor Rf

  30. Dissection Instruments • Tools used to perform dissection of Tissues/ organisms • Dissecting scope sometimes used

  31. Micro dissection Instruments • Tools used to perform dissection of individual cells or the removal of cell organelles. • Most commonly used with the Electron microscope (greatest magnification).

  32. Staining Techniques • The use of solutions to make it more possible for an observer to see structures of a cell easier under a microscope 1. Iodine- used primarily to stain nucleus of animal cell. 2. Methylene blue- used primarily to view chloroplasts and the cell wall of a plant cell.

  33. Staining Techniques

  34. Indicators • Designed to detect specific materials INDICATORS DETECTS • Iodine/ Lugols solution  starch • Benedicts solution  Glucose • Biuret solution  proteins • Bromomethyl blue  carbon dioxide and Lime water • Litmus paper  acids or bases

  35. SI: System International, The Metric System • The most common system of measurement used by scientists is the metric system. The metric system is based on the unit 10. (Based on multiples of ten) The metric system has many basic units for its’ measurements. (Length = is a meter, etc.)

  36. SI: System International, The Metric System • Think of the basic units as Suffixes. *Meter, Gram. Second, Liter, Degree Celsius, • Think of the abbreviations as prefixes: • Kilo =1000 • Centi = 100th • Milli = 1000th

  37. ALLLL DONE!