Chapter 3Chapter 3
Biological tools Biological tools and and
InstrumentationInstrumentation
Taxonomic Taxonomic (Dichotomous) (Dichotomous)
KeyKey1a. Is the organism unicellular -go to 1a. Is the organism unicellular -go to step 2step 2
1b. Is the organism multicellular -go to 1b. Is the organism multicellular -go to step 3step 3
2a. Is the organism prokaryotic -kingdom 2a. Is the organism prokaryotic -kingdom MoneraMonera
2b. Is the organism eukaryotic -Kingdom 2b. Is the organism eukaryotic -Kingdom ProtistaProtista
Scientific MethodScientific MethodAn systematic approach to problems An systematic approach to problems
solving:solving: 1. Defining the problem1. Defining the problem
“ “ By doing Background ResearchBy doing Background Research”” 2. Formulating a hypothesis2. Formulating a hypothesis 3. Testing the hypothesis3. Testing the hypothesis 4. Recording observations4. Recording observations
5. State a conclusion5. State a conclusion
Three types of ExperimentsThree types of Experiments
1.1. ****Controlled ****Controlled Experiments**** Experiments****
2.2. Trial and Error Trial and Error
3.3. Open EndedOpen Ended
Defining the problemDefining the problem ““By doing Background By doing Background
ResearchResearch””HypothesisHypothesis – –ExperimentationExperimentation
1. 1. Control groupControl group (no change) (no change)
2. 2. Experimental groupExperimental group (make (make change)change)
control =control = variable =variable =
** State a conclusionState a conclusion
Defining the problemDefining the problem
HypothesisHypothesis – –a possible explanation for a possible explanation for events.events.
ExperimentationExperimentation (Testing the (Testing the hypothesis)hypothesis)
1. 1. Control groupControl group (no change) a source (no change) a source for for comparison comparison
2. 2. Experimental groupExperimental group (contains 2 (contains 2 variables)variables)
*Independent Variable (Manipulated) *Independent Variable (Manipulated) **Dependent Variable (responding) Dependent Variable (responding)
#’s#’sState a conclusionState a conclusion
TheoryTheory- explanation - explanation supported by experimental supported by experimental evidence.evidence.
Scientific LawScientific Law- is a - is a statement that describes statement that describes some aspect of a phenomena some aspect of a phenomena that is always true.that is always true.
MicroscopesMicroscopes1.Simple Microscopes2.Dissecting microscope3.Compound Microscope4.Phase Contrast Microscope5.Electron Microscope
• SEM• TEM
InstrumentationInstrumentationA. A. Simple microscope Simple microscope - magnifying - magnifying glassglass
1 –uses 1 lens1 –uses 1 lens B. B. Compound light microscopeCompound light microscope 1- uses two lenses1- uses two lenses a) ocular (eyepiece) 10xa) ocular (eyepiece) 10x b) objective b) objective - low power, 10x- low power, 10x - high power, 40x- high power, 40x
Compound Light Compound Light MicroscopeMicroscope
Magnifies 50x Magnifies 50x to 500x the to 500x the original sizeoriginal size
Used to look at Used to look at the whole cell the whole cell and organelles and organelles like the nucleus like the nucleus and chloroplastand chloroplast
Adjustments:Adjustments:
Coarse adjustmentCoarse adjustment – used to – used to focus under low power focus under low power
Fine adjustmentFine adjustment – used to – used to focus under high power focus under high power ONLYONLY
Used to “fine tune” the Used to “fine tune” the imageimage
Total Total MagnificationMagnification::
Magnifications: Magnifications: ocularocular = 10x = 10x
low power objective = 10xlow power objective = 10x
high power objective = 40xhigh power objective = 40x
Formula:Formula:
ocularocular x objective = total magnification x objective = total magnification
High power magnification is? High power magnification is?
1010 x 40 = 400x x 40 = 400x
Imagery:Imagery:When placing a slide under the When placing a slide under the
objective lens, the image objective lens, the image appears:appears:
-magnified-magnified-upside down-upside down-inverted (backwards).-inverted (backwards).
ExEx: e : e F F
Imagery:Imagery:(as seen under low (as seen under low
power)power)e e
F F
FF ee
Imagery:Imagery:Field of View (fov)Field of View (fov)
Scanner fov = 4000 um
F FLow fov=2000 um
What is the size of the “F”?
Imagery:Imagery:Field of View (fov)Field of View (fov)
FF 4000 um
Electron MicroscopesElectron MicroscopesAdvanced Advanced
microscope microscope magnifies 250,000x magnifies 250,000x
the original sizethe original sizeAllows for detailed Allows for detailed
observation of observation of smallsmall organellesorganelles within the cell. within the cell. Like the Like the
mitochondria and mitochondria and lysosomes.lysosomes.
Dissecting MicroscopeDissecting MicroscopeGives the viewer a Gives the viewer a
3-D image of the 3-D image of the organismorganism
Used in the Used in the dissection of small dissection of small animals and plantsanimals and plants
Low magnificationLow magnification1-10x1-10x
Other Biological ToolsOther Biological ToolsSeparation TechniquesSeparation Techniques
CentrifugeCentrifuge ChromatographyChromatography Gel electrophoresisGel electrophoresis Dialysis TubingDialysis Tubing
Dissection and micro dissection Dissection and micro dissection instrumentsinstruments
StainsStains IndicatorsIndicatorsSystem InternationalSystem International
Separation TechniquesSeparation Techniques
Centrifuge:Centrifuge: Separates of solutions based on each Separates of solutions based on each parts densityparts density
Chromatography:Chromatography: Separates Solute Separates Solute from Solvent from Solvent
Electrophoresis:Electrophoresis: separates macromolecules-separates macromolecules-either nucleic acids or proteins-on the basis of size, either nucleic acids or proteins-on the basis of size, electric charge, and other physical properties electric charge, and other physical properties
Dialysis TubingDialysis Tubing
CentrifugeCentrifuge
Separates of Separates of solutions based solutions based on each parts on each parts densitydensity
Gel ElectrophoresisGel Electrophoresis
ElectrophoresisElectrophoresis
Opposite charges at each side Opposite charges at each side causing sample to movecausing sample to moveSmaller DNA fragments move Smaller DNA fragments move faster and furtherfaster and further
IV. ElectrophoresisIV. Electrophoresis
Electrophoresis:Electrophoresis: separates nucleic acids or proteins-on the basis separates nucleic acids or proteins-on the basis
of size, electric charge, and other physical of size, electric charge, and other physical properties properties
Chromatography Chromatography Separates Solute from Solvent Separates Solute from Solvent
Distance traveled by the solute (ink)Distance traveled by the solute (ink)
Distance traveled by the solvent (liquid)Distance traveled by the solvent (liquid)Rf=Rf=
Thin-Layer Chromatography Thin-Layer Chromatography (TLC)(TLC)
ChromatographyChromatography
Retention Factor RfRetention Factor Rf
Dissection InstrumentsDissection InstrumentsTools used to perform Tools used to perform
dissection of Tissues/ dissection of Tissues/ organisms organisms
Dissecting scope Dissecting scope sometimes usedsometimes used
Micro dissection Micro dissection InstrumentsInstruments
Tools used to perform Tools used to perform dissection of individual dissection of individual cells or the removal of cell cells or the removal of cell organelles.organelles.
Most commonly used with Most commonly used with the Electron microscope the Electron microscope (greatest magnification).(greatest magnification).
Staining Staining TechniquesTechniquesThe use of solutions to make it more The use of solutions to make it more
possible for an observer to see possible for an observer to see structures of a cell easier under a structures of a cell easier under a microscopemicroscope
1. 1. IodineIodine- used primarily to stain - used primarily to stain nucleus of animal cell.nucleus of animal cell.
2. 2. Methylene blueMethylene blue- used primarily to - used primarily to view chloroplasts and the cell wall of a view chloroplasts and the cell wall of a plant cellplant cell..
Staining Staining TechniquesTechniques
IndicatorsIndicatorsDesigned to detect specific materialsDesigned to detect specific materials
INDICATORSINDICATORS DETECTS DETECTS Iodine/ Lugols solution Iodine/ Lugols solution starchstarchBenedicts solution Benedicts solution GlucoseGlucoseBiuret solutionBiuret solution proteins proteins Bromomethyl blue Bromomethyl blue carbon dioxidecarbon dioxide
and Lime waterand Lime waterLitmus paperLitmus paper acids or bases acids or bases
SI: System International, SI: System International, The Metric SystemThe Metric System
The most common system of The most common system of measurement used by scientists is measurement used by scientists is the metric system. The metric the metric system. The metric system is based on the unit 10. system is based on the unit 10. (Based on multiples of ten) The (Based on multiples of ten) The metric system has many basic units metric system has many basic units for its’ measurements. (Length = is a for its’ measurements. (Length = is a meter, etc.) meter, etc.)
SI: System International, SI: System International, The Metric SystemThe Metric System
Think of the basic units as Suffixes. Think of the basic units as Suffixes.
*Meter, Gram. Second, Liter, Degree *Meter, Gram. Second, Liter, Degree Celsius,Celsius,
Think of the abbreviations as prefixes:Think of the abbreviations as prefixes:Kilo =1000Kilo =1000Centi = 100Centi = 100thth
Milli = 1000thMilli = 1000th
ALLLL DONE!ALLLL DONE!