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BIOL 251 001-2
CHP. 3
| Question | Answer |
|---|---|
| THE 5 I'S CULTURING MICROBES | 1. INOCULATION 2. INCUBATION 3. ISOLATION 4. INSPECTION 5. IDENTIFICATION |
| INOCULATION | INTRODUCTION OF A SAMPLE INTO A CONTAINER OF MEDIA |
| INCUBATION | UNDER CONDITIONS THAT ALLOW GROWTH |
| ISOLATION | 1. SEPARATING ONE SPECIES FROM ANOTHER 2. IF AN INDIVIDUAL BACTERIAL CELL IS SEPARATED FROM OTHER CELLS & HAS SPACE ON A NUTRIENT SURFACE, IT WILL GROW A MOUND OF CELL - COLONY |
| INSPECTION | TAKE SMEARS & PREPARE A SLIDE TO VIEW |
| IDENTIFICATION | |
| COLONY | A COLONY CONSISTS OF ONE SPECIES |
| ISOLATION TECHNIQUE | 1. MIXTURE OF CELLS IN SAMPLES 2. SEPARATION OF CELLS BY SPREADING OR DILUTION ON AGAR MEDIUM 3. INCUBATION ALLOWS GROWTH INCREASES THE # OF CELLS 4. MICROBES BECOME VISIBLE AS ISOLATED COLONIES CONTAINING MILLIONS OF CELLS |
| MEDIA | PROVIDING NUTRIENTS IN THE LABORATORY |
| MOST COMMONLY USED MEDIA | 1. NUTRIENT BROTH 2. NUTRIENT AGAR |
| NUTRIENT BROTH | LIQUID MEDIUM CONTAINING BEEF EXTRACT & PEPTONE |
| NUTRIENT AGAR | SOLID MEDIA CONTAINING BEEF EXTRACT, PEPTONE & AGAR |
| AGAR | 1. A COMPLEX POLYSACCHARIDE ISOLATED FROM RED ALGAE 2. SOLID AT ROOM TEMP, LIQUEFIES AT BOILING (100C), DOESN'T RESOLIDIFY UNTIL COOLS TO 42C 3. PROVIDES FRAMEWORK TO HOLD MOISTURE & NUTRIENTS NOT DIGESTIBLE FOR MOST MICROBES |
| CLASSIFICATION OF MEDIA | 1. PHYSICAL STATE - LIQUID, SEMI-SOLID, SOLID 2. CHEMICAL COMPOSITION: SYNTHETIC OR NONSYNTHETIC 3. FUNCTION TYPE: GENERAL PURPOSE, ENRICHED, SELECTIVE DIFFERENTIAL, ANAEROBIC (REDUCING) |
| SYNTHETIC MEDIUM | CONTAINS PURE ORGANIC & INORGANIC COMPOUNDS IN AN EXACT CHEMICAL FORMULA |
| NONSYNTHETIC (COMPLEX) MEDIUM | CONTAINS AT LEAST ONE INGREDIENT THAT IS NOT CHEMICALLY DEFINABLE |
| GENERAL PURPOSE MEDIA | GROWS A BROAD RANGE OF MICROBES, USUALLY NONSYNTHETIC |
| ENRICHED MEDIA | CONTAINS COMPLEX ORGANIC SUBSTANCES SUCH AS BLOOD, SERUM, HEMOGLOBIN OR SPECIAL GROWTH FACTORS REQUIRED BY FASTIDIOUS MICROBES |
| SELECTIVE MEDIA | CONTAINS ONE OR MORE AGENTS THAT INHIBIT GROWTH OF SOME MICROBES AND ENCOURAGE GROWTH OF THE DESIRED MICROBES |
| DIFFERENTIAL MEDIA | ALLOW GROWTH OF SEVERAL TYPES OF MICROBES AND DISPLAYS VISIBLE DIFFERENCES AMONG DESIRED AND UNDESIRED MICROBES |
| REDUCING MEDIUM | 1. CONTAINS A SUBSTANCE THAT ABSORBS OXYGEN OR SLOWS PENETRATION OF OXYGEN INTO MEDIUM 2. USED FOR GROWING ANAEROBIC BACTERIA |
| CARBOHYDRATE FERMENTATION MEDIUM | CONTAINS SUGARS THAT CAN BE FERMENTED, CONVERTED TO ACIDS, AND A (LOW) pH INDICATOR TO SHOW THE REACTION; BASIS FOR IDENTIFYING BACTERIA & FUNGI (THAT LIKE THE ACIDIC ENVIRONMENT) |
| MAGNIFICATION | THE ABILITY TO ENLARGE AN OBJECT |
| RESOLVING POWER | THE ABILITY TO SHOW DETAILS |
| Bright-field | most widely used, specimen is darker than surrounding field |
| Dark-field | brightly illuminated specimens surrounded by dark field |
| Phase- Contrast | transforms subtle changes in light waves passing through the specimen into differences in light intensity, best for observing intracellular structures |
| Fluorescence Microscope | 1. Modified compound microscope with an ultraviolet radiation source and a filter that protects the viewer’s eye 2. Uses dyes that emit visible light when bombarded with shorter uv rays. 3. Useful in diagnosing infections |
| Electron microscopy | 1. Forms image w/ a beam of electrons that can be made to travel in wavelike patterns when accelerated to high speeds 2. Electron waves are 100,000X shorter than the waves of visible light 3. Electrons have tremendous power to resolve tiny structures |
| Types of electron microscopes | 1. Transmission electron microscopes 2. scanning electron microscopes |
| Transmission electron microscopes | transmits electrons through the specimen; darker areas represent thicker, denser parts and lighter areas indicate more transparent, less dense parts |
| Scanning electron microscopes | provides detailed three-dimensional view. SEM bombards surface of a whole, metal-coated specimen with electrons while scanning back and forth over it. |
| specimen preparation | 1. wet mounts & hanging drop mounts 2. Fixed mounts 3. Smear |
| Wet mounts & hanging drop mounts | allow examination of characteristics of live cells: motility, shape & arrangement |
| Fixed mounts | made by drying & heating a film of specimen |
| Smear | A stained using dyes to permit visualization of cells or cell parts |
| Staining | 1. Cationic dyes 2. Anionic dyes 3. Positive staining 4. Negative staining 5. simple stains 6. differential stains 7. Special stains |
| Cationic dyes | Basic, with positive charges on the chromophore |
| Anionic dyes | Acidic, with negatice charges on the chromophore |
| Positive Staining | Surfaces of microbes are negatively charged and attract basic dyes |
| Negative Staining | Microbe repels dye & stains the background |
| Simple Stains | One dye is used |
| Differential stains | Surfaces of microbes are negatively charged and attract basic dyes |
| Special stains | Capsule & flagellar stains |
| Resolution | The Capacity to distinguish separate of 2 adjacent object |
Created by:
ph4tm4m4
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