Micro Chapter 3: Tools Of The Lab

Front 1

The 5 I's

Back 1

inoculation
incubation,
isolation
inspection
identification

Front 2

Inoculation

Back 2

Producing a culture.
Introduce a tiny sample (the inoculums) into a container of nutrient medium

Front 3

Isolation

Back 3

separating one species from another

Front 4

Streak plate method

Back 4

Small droplet of culture or sample spread over surface of the medium with an inoculating loop.

Uses a pattern that thins out the sample and separates the cells.

Purpose is to separate.

Front 5

Loop dialation method
(serial dilution method)

Back 5

Sample inoculated serially in to a series of liquid agar tubes to dilute the number of cells in each successive tubes.

Makes it workable w/reasonable amount of colonies

Front 6

Spread plate method

Back 6

small volume of LIQUID, diluted sample pipette on to surface of the medium and spread around evenly by a sterile spreading tool.

Not as "good" as serial dilution method.

Front 7

Three categories of Media Classification

Back 7

Physical State
Chemical Composition
Functional Type

Front 8

Physical State

Back 8

1) Liquid (broths, milks, infusions)
2) Semisolid (clot-like at RT)
3) Solid (can be converted to a liquid; firm surface where colonies can grow)
4) Solid (cannot be liquidfied)

Front 9

Chemical Composition

Back 9

1) Synthetic- you added chemicals, but it is defined. You know exactly what you added.

2) Nonsynthetic- complex; not chemically defined. ex: added yeast- don't know exactly what is in yeast.

Front 10

Synthetic Media

Back 10

compositions are precisely chemically defined

Front 11

Nonsynthetic (complex) media

Back 11

If even just one component is not chemically definable, than it is complex.

Front 12

Functional Type:
General Purpose Media

Back 12

-to grow as broad a spectrum of microbes as possible
-Usually nonsynthetic
-Contain a mixture of nutrients to support a variety of microbes
-you will exclude MOs that like low-nutrient concentration

Front 13

Functional Type:
Enriched Media

Back 13

-Add components to help organism grow
-contain complex organic substances (for example blood, serum, growth factors) to support the growth of fastidious bacteria. Examples: blood agar, Thayer-Martin medium (chocolate agar)

Front 14

Functional Type:
Selective media
(Defined, synthetic)

Back 14

To particularly separate and grow a particular organism and not others.

Front 15

Functional Type:
Differential media
(undefined, complex)

Back 15

Organisms will look different (color), but it will not stop any organism from growing.

Front 16

Incubation

Back 16

an inoculated sample is placed in an incubator to encourage growth.

Can change temp, atmosphere, etc

Front 17

Inspection and Identification

Back 17

Using appearance as well as metabolism (biochemical tests) and sometimes genetic analysis or immunologic testing to identify the organisms in a culture.

Front 18

Total magnification

Back 18

ocular power x objective power

Front 19

Objective lens

Back 19

forms real image

Front 20

Ocular lens

Back 20

forms the virtual image

Front 21

Resolution

Back 21

The ability to distinguish two adjacent objects or points from one another. Dependant on wavelength of light.

Also known as resolving power

Front 22

Magnification and resolution

Back 22

increased magnification decreases the resolution

Front 23

Bright-Field Microscopy

Back 23

-Most widely used
-Forms its image when light is transmitted through the specimen
-The specimen produces an image that is darker than the surrounding illuminated field
-Can be used with live, unstained and preserved, stain specimens

Front 24

Dark-Field Microscopy

Back 24

Adds a little more contrast. Specimen is light, with dark field around it.

Front 25

Phase Contrast Microscopy

Back 25

Use with a living specimen.

Allows differentiation of internal components of live, unstained cells

Front 26

Interference Microscopy

Back 26

Allows for detailed view of live, unstained specimens

The image is colorful and 3D

Front 27

Fluorescence Microscopy

Back 27

Used for diagnosing and counting

Front 28

Confocal Microscopy

Back 28

Uses a laser beam. Allows for viewing cells a a higher magnification.

Used on fluorscent stained specimens

Front 29

Electron Microscopy

Back 29

Allows scientists to view the finest structure of cells.

Two types: Transmission electron (TEM) and Scanning electron (SEM)

Front 30

Transmission Electron Microscope (TEM)

Back 30

Better for internal visualization

can see cross sections of specimens

Front 31

Scanning Electron Microscope (SEM)

Back 31

Better for external/surface visualization

Front 32

Wet mounts on a slide

Back 32

Cells suspended in fluid, a drop or two of the culture is then placed on a slide and overlaid with a cover glass

Front 33

Hanging drop mount

Back 33

Uses a depression slide, Vaseline, and coverslip. Better for bigger organisms.

Front 34

Smear technique

Back 34

smear organism on slide, usually use heat to adhere

Front 35

Stain technique

Back 35

stain penetrates for contrast

Front 36

Positive stain

Back 36

cells take on stain color

Front 37

Negative Stain

Back 37

surroundings take on stain, creating a silhouette of the specimen.

Front 38

Simple stains

Back 38

require only a single dye; everything is the same color.

Front 39

Differential stains

Back 39

Two differently colored dyes. Things look different (pink cells, purple cells).
More steps involved.

Front 40

Gram Staining

Back 40

Diagnostic staining technique.
Positive= purple
Pink= negative

Front 41

Acid-Fast Staining

Back 41

Useful for specific bacteria

Diagnostic stain

Front 42

Endospore Staining

Back 42

Dye is forced by heat into resistant bodies called spores or endospores.

More difficult to do.