Ch 2

what are the building blocks of proteins

amino acids

building blocks of carbohydrates

glucose

building blocks of nucleic acids

nucleotides

building blocks of lipids

fatty acids

what in the cell membrane made of

phospholipids

small RNA micro advancement

first identified in the 2000s, and appeared to play a role in regulating what happens in cells

PCR micro advancement

a technique developed in the 1980s to make mass copies of DNA for study

restriction enzymes micro advancement

discovered is 1970s and called “molecular scissors” that cut DNA ina controlled manner

Human Microbiome Project (HMP) micro advancement

in the 2010s and beyond, it provides detailed info about the microorganisms that live on or in our bodies

prokaryotic microbe that lakcs a true membrane-bound nucleus, but has genome DNA as a circular chromosome suspended in the cytoplasm

bacterium

acellular infectious protein

prion

process in which humans manipulate the genes of microorganisms

genetic engineering

five basic techniques to manipulate, grow, examine, and characterize microorganisms in the lab

inoculation, incubation, isolation, inspection, identification

inoculation means

producing a culture

what is a culture

a cultivation of microorganisms or any growth that appears on a medium after incubation

medium plural

media

medium

a nutrient containing enviorment where microbes can multiply

bacteria are festidius meaning

they are picky about what they eat

inoculation

introduction of microbes into media for culture

sterile

free of microbes, especially in a medical setting

being sterile is a requirement for

any instrument used for sampling and inoculation

incubation

control asmospheric gases to allow microbial growth on media

usual temperatures for incubation

20 to 45 degrees celsius

what atmospheric gases do incubators control

carbon and oxygen

turbidity

cloudiness

what type of microbial growth do you see in liquid media

turbidity, sediment, scum, and color changes

what type of microbial growth do yoou see on solid media

wisible piled up cells or colonies

isolation

a small numbe rof cells must be inoculated into a relatively large volume or expansive area to encourage growth

do broths isolate colonies

no, because they are a liquid

what is needed to isolate a colony

firm surfaces or petri dish, inoculating tools (ex: inoculating loop)

types of techniques for obtaining isolated colonies and pure cultures

streak plate or pour plate

pure culture

container of medium with only a single known species or type of microorganism

anexic

free of other living things except for the one being studied

subculture

a second-level culture from a well-isolated colony, additional culturing from original culture

mized culture

a container that holds two or more identified, easily differentiated species of microorganisms

contaminated culture

a culture that was once ure or mixed but now contain contaminants or unwnated microbes of uncertain identity

fundamental chemical chracteristics gotten from biochemical testing

nutrient requirements, products given off during growth, presence of enzymes, and mechanisms for deriving energy

biochemical tests are a part of

inspection and identification

what products are given off of bacteria

acids, gases, sugars, alcohols

mechanisms of deriving energy

aerobic, anerobic

genetypic testing

detects microbes based on their DNA

immunologic testing

testing the isolate against known antibodies

analytical or diagnostic tools

genotypic testing, immunological testing, and inoculation of suitable lab animals

microbial profiles are determined through combining

phenotypic testing, genotypic testing, immunologic tesring, macroscopic analysis, and microscopic analysis

genotypic testing

molecular genetics

microbes range in size from

20nm (virus) to 1mm (bacillus megaterium)

name in order from smallest to largest:

bacillus, chlamydia, strep, protozoa, bacteria, mycoplasma, viruses, staph, clostridium and rickettsia

viruses, chlamydia, rickettsia, mycoplasma, bacteria, staph, strep, bacillus, clostridium, protozoa

microscope we use in micro

compoud light microscope

wavelength

distance from one part of a wave to the next part

visible light ranges from

400 nm to 700 nm

the range of light is approx. the same size as a lot of bacteria especially the

400 nm region

the best resolution for seeing most bacteria is the use of

blue light, using a blue filter

resolving power

ability to distinguish 2 or more objects from each other

why is resolving power important

you would just see what looks like a mass of cells

is blue or red light better for looking at resolving bacteria

blue light

reflection

light waves bounce off of an object

transmission

waves pass through an object

bacteria don’t have the ability for light to bouce back because

the light can’t go through them

absorption

wave energy gained by an object is given off at a different energy level

example of absorption

fluorescence, being able to see things that glow

refraction

bending of waves as they pass through two different media

example of refraction

using immersion oil to increase resolution by reducing the amount of refraction going from air to glass to help better see the bacteria

diffraction

waves bend around a barrier or object

contrast

the ability to distinguish objects from each other and their background

compound light microscope

common in micro lab, uses bright-field or dark-field

bright field

background is illuminated, objects look darker

cons to bright field

poor contrast without staining cells

pros to bright field

good resolution and good contrast when dyes are used to stain bacterial cells

dark field

background is dark, objects look brighter

pros to dark field

good contrast

cons to dark field

limited differential properties

phase contrast microscope

has a dual beam for viewing objects in 3D with increased depth perception

dual beam

has two beams of light to get the 3D effect

fluorescent microscopy

uses UV light to illuminate the bacteria

fluor-chromes

addition of fluorescent chemicals to increase contrast and specigicity under light

fluorescent antibody staining

uses fluorescent-labeled antibodies to specifically stain a bacterial cell or other substances/objects

elecctron microscopy (EM)

uses electron beams to see things smaller than visible light wavelengths

what can an electron microscope see

viruses, proteins, small organelles (ex: ribosomes), and atoms

what size objects can an electron microscope see

usually object 0.1 nm to 10nm in size, even larger with SEM or TEM

TEM

transmission electron microscopy

transmission electron microscopy

electrons pass through something

SEM

scanning electron microscopy

scanning electron microscopy

seeing surface features

two types of scanning-tunneling and atomic force microscopy

visualizing molecules and atoms and manipulating (moving) atoms

types of light microscope speciman preparation

wet mounts and smears

wet mounts

allows one to view living cell

what are you sble to see in a wet mount speciman

shape, movement (flagella/cilia), internal and external structures

smears

suspension of cells smeared on a glass microscope slide

how are smeared bacteria put onto slide

usually heat-fixed or air dryed so cells attach and don’t wash off when staining

why do the cells need to stick to slide

theres lots of rinsing in smears so the cells need to stick to the slide

what happens to cells once they are smeared onto slide

they die (no longer pathogenic) so you can’t see movement or other viable charcteristics

another name for acidic dyes

negative stains or dyes

acidic dyes

stain acidophillic structures

acidophilic structures

acid loving

examples of acidic dyes

nigrosin and congo red

how do acidic dyes work

the pigmented part of the molecule is negtively charged so the dyes will stick to positively charged substances and structures

is a cytoplasmic protein is positively charged what will the acidic dyes stick to

the dyes are negative so it will stick to the protein

if bacteria cells are negatively charged what will the acidic dyes stick to

the dyes are negative so it will not stick to the bacteris but instead the background surrounding the bacteria

mpost bacterial dyes are

basic ones

basic dyes

stain basophilic structures and cells