Experiments 3,4 & 5: Simple & Gram Stain, Pure Culture Isolation & Transfer Method, And Differential Staining

Front 1

Staining techniques are used to ______ the specimen

Back 1

color

Front 2

INFO ONLY: Most microorganisms are transparent & by staining we can give a color contrast between

Back 2

the cells and the background to better view/ study the cells.

Front 3

What does morphologies mean?

Back 3

Shapes

Front 4

coccobacilli shape? Pleomorphic? Staphylo? Strepto? Sarcina?

Back 4

Short rods; multiple shapes; clusters; chains; two quartets of cell that form a cube shaped

Front 5

What is the 1st step in preparing a specimen to be stained?

Back 5

preparing a smear

Front 6

In this technique a sample of bacteria is applied to a slide and allowed to dry? A similar technique requires passing the slide through the flame a few times?

Back 6

fixation; heat fixation

Front 7

Which culture requires a drop of water? This culture is in a suspension?

Back 7

Agar culture; broth culture

Front 8

In this stain, it stains the specimen but not the background with the light passing through the background and the colored cells are easily visible?

Back 8

Direct stain

Front 9

This technique stains the background, but not the specimen and the background will be colored and the light will pass through the cells?

Back 9

Negative stain

Front 10

Stains a bacterial smear with a single dye, cells are colored on an unstained background?

Back 10

Simple stain

Front 11

* This type of stain is primarily used to determine the morphology, size and arrangement of the bacterial cells?

Back 11

Simple stain

Front 12

A stain the uses at least three chemical reagents on a smear?

Back 12

Differential stain

Front 13

This stain generally gives you more information on a bacterium compared to a simple stain?

Back 13

Differential stain

Front 14

The most common differential stain is the ______ stain. Who developed this stain?

Back 14

Gram; Hans Gram

Front 15

This stain is used to differentiate bacteria based on their cell wall composition?

Back 15

Gram stain

Front 16

What are the 4 classes of reagents used in differential staining? What are the names of these reagents?

Back 16

Primary stain, decolorized, mordant and counter stain; Crystal violet, ethanol, iodine and safranin.

Front 17

Staphylococcus epidermidis is a gram _______ cocci bacteria? Escherichia Coli is a gram ______ rod bacteria used in the simple and gram stain experiments.

Back 17

Positive; negative

Front 18

If the colors are switched for these two bacteria then what does this possibly mean?

Back 18

Over/under-decolorized

Front 19

Contains only one type of microorganism and is important in the ID + classification of bacteria?

Back 19

Pure culture

Front 20

Transferring bacteria from one medium to another? This technique requires sterile equipment and sterile conditions into a medium?

Back 20

Subculturing; aseptic transfer

Front 21

This technique separates mixed cultures of bacteria into individual colonies? What does this technique do to the quantity of organisms on the plate?

Back 21

Streak plate technique; Reduces

Front 22

(INFO only) The streak plate technique is important to allow the different species of bacteria in the culture to be spread far enough apart on the agar so

Back 22

that the individual cells can multiply without touching.

Front 23

What are the 7 criteria used when describing a colony's morphology.

Back 23

Size, pigment, optical property, surface, form, margin, and elevation. (POSSEM F)

Front 24

Three additional differential stains besides gram that were developed are what?

Back 24

Capsule, endospore and acid-fast stain

Front 25

The capsule in a bacterium is resistant to most stains and ________-soluble. In the experiment the capsule will seen as a halo because of its resistance while the rest of the bacterium is stained purple or red?

Back 25

water; purple

Front 26

What staining method is used for the encapsulated bacteria?

Back 26

Anthony method

Front 27

An inactive cell type that is highly resistant to harsh environments? These cell type resist stains better than vegetative cells, yes or no?

Back 27

Endospores; Yes they are very resistant

Front 28

What method is used to stain the endospores? What technique is used to help drive the primary stain into the spore? What counterstain is used for the vegetative cells?

Back 28

Malachite green method; Heat( steaming); Safranin(pink)

Front 29

These organisms are able to resist the decolorizer acid-alcohol are called? What method is used to stain these bacteria?

Back 29

Acid-fast; Ziehle-Neelsen method

Front 30

Why are these bacteria successful at preventing decolorizing reagents? What is the primary stain used in these stains?

Back 30

Because these bacteria have a waxy coat around their cells which hinders the entry of stains; Carbol fuchsin(red)

Front 31

In the stain non-acidfast organisms will stain what color? Acid-fast organisms will stain which color?

Back 31

Methylene blue; Red

Front 32

What bacteria was used in the capsule stain? Capsule stain is a _______stain where the background and bacterial cells will appear purple.

Back 32

Klebsiella pneumoniae; negative

Front 33

What are the 2 bacterium used in the endospore stain? Which bacterium should have more spores?

Back 33

Young and old Bacillus subtilis; older B. bubtilis

Front 34

What are the 2 bacteria used in acid-fast? What color will these 2 bacterium ensue?

Back 34

M. smegmatis and S. aureus; S. aureus is a blue cocci and M. smegmatis is a red rods