Final Chapter 3 - Linda Jane (staining)

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few, Bacillus, when they are stressed

CHECK THIS ONE>>>>>>>many?/ few? types of bacteria produce spores including ___ genus. why?

to enhance contrast to observe with microscope, as most of them have transparent cytoplasm

Why do we do Simple Staining to identify microorganisms?

most cells are - so we use + stains

most bacteria are -?/ +? therefore we use -?/ +? to make a simple stain

kills the bacteria,
makes specimen stick to slide,
coagulates their proteins for better, staining.

Why is heat fixing done when staining? (give three reasons)

water

when making a simple stain: after applying crystal violet, safranin or methylene blue stain, rinse with ___

methylene blue, safranin, crystal violet

what three stains are most often used for simple staining?

nigrosin,
acidic

___ are/is example(s) of negative stain(s)

methylene blue, safranin, crystal violet

___ are/is example(s) of positive stains

negative, stain

when making a simple?/negative? stain, the first step is to put the microorganism in a drop of ___ and smear it on a slide using a slide spreader

wait for slide to air fix (dry) and observe

in negative staining: after making a smear from an inoculated drop of stain on a slide what happens next?

cells are too delicate to withstand heat fixing

why do we use a negative stain technique?

negative staining

which kind of staining technique colors the background and not the cells?

nigrosin (if the cells are negatively charged), acidic

what stain might we use in a negative stain and is it acidic or basic stain

negative staining

which kind of staining technique do we use to observe halo?

purple

what color is nigrosin stain?

differential

to tell if a cell is gram positive or gram negative we use ___ stain technique (NOT looking for the word 'gram' here)

differential

gram staining is a type of ___ staining

if the cell is gram - or + AND the morphology, size and arrangement

gram staining tells us two main things: ___

gram

SIMPLE?/ GRAM?/ NEGATIVE?/ ACID-FAST? staining is typically the FIRST staining tecnnique you would apply to a cell.

(1) heat fix microorganism
(2) primary stain (with basic (+) stain such as crystal violet)
(3) apply mordent (such as iodine to enhance violet staining)
(4) decolorize (with ethanol for example)
(4) counter stain (with safrinin)

the general steps in gram staining are:

negative, because the ethanol melts away the lipid layer and the dye leaks out the thinner peptoglycan wall.

gram + or - cells loose the purple color when decolorizing in gram staining. Why?

crystal violet

what is the first dye you add in gram staining?

do

in simple staining DO?/ DO NOT? heat fix

do not

in negative staining DO?/ DO NOT? heat fix

after, heat

in simple staining apply the stain before?/ after air?/ heat? fixing

acidic

when making a negative stain, the microorganism is added to a drop of basic stain?, acidic stain?/ water?

(1) heat fix microorganism
(2) cover slide with bibulous paper and apply carbolfuchsin stain and steam for 5 minutes
(3) rinse with water and then DECOLORIZE with ethanol
(4) counter stain with methylene blue, rinse, blot and observe

the general steps in acid-fast staining are:

simple stain:
(1) heat fix
(2) apply a stain (usually positive for example methylene blue, safranin, crystal violet)
(2) wait 1 min and rinse with water, blot and observe

the general steps in simple staining are:

negative stain:
(1) add drop of nigrosin stain to slide
(2) inoculate stain on slide
(3) spread inoculated stain on slide and AIR dry, observe

the general steps in negative staining are:

acid-fast stain:
(1) heat fix
(2) carbolfuchsin stain, 5
(3) DECOLORIZE with ethanol
(4) methylene blue

in acid-fast staining:
in ___ staining technique:
(1) ___ microorganism
(2) cover slide with bibulous paper and apply ___ stain and steam for ___ minutes
(3) carefully remove paper, rinse with water and then ___
(4) counter stain with ___, rinse, blot and observe

(1) add congo red stain to serum
(2) smear with spreader slide
(3) AIR dry
(4) flood with maneval's stain, rinse, blot and observe

the general steps in capsule staining are:

capsule

the general steps in ___ staining are:
(1) add congo red stain to serum
(2) smear with spreader slide
(3) AIR dry
(4) flood with maneval's stain, rinse, blot and observe

capsule and negative

you do NOT heat fix in which staining techniques?

(1) heat fix microorganism on slide
(2) cover with bibulous paper and apply malachite green stain, steam for 5 minutes
(3)carefully remove paper, rinse with water and counter stain with safranin stain, rinse, blot and observe

the general steps in endospore staining are

endospore
1) heat fix
(2) malachite green
(3) safranin stain

the general steps in ___ staining are
(1) ___ microorganism on slide
(2) cover with bibulous paper and apply ___ stain, steam for 5 minutes
(3) carefully remove paper, rinse with water and counter stain with ___ stain, rinse, blot and observe

because bacteria with a high content of mycolic acid (a wazy substance in their cell walls) are resistant to ethanol and do NOT decolorize in differential staining when ethanol is applied.
These bacteria might be of type Mycobacterium (which are highly pathogenic) or some other cell with mycolic acid.

why do we apply an acid-fast staining tecnhique?

"mother" cells are red and spore are green with malachite green

what is the expected result of endospore staining?

are, because they have keratin in their outer coating

endospores are?/ are not resistant to heat fixing and staining. Why?

endospore, it is water soluable and can be forced into the spore by steaming

malachite green stain is used in ___ staining. Why?

(1) heat fix microorganism onto slide
(2) cover with bibulous paper and apply malachite green stain, steam for 5 minutes
(3) carefully remove paper, rinse with WATER to decolorize
(4) counterstain with safranin, leave fore 1 minute, rinse an, blot and observe

the general steps in endospore staining are:

a negative stain with halo effect (no color) in capsule.

in capsule staining what result do we expect

negative

do we use a negative or positive stain in capsule staining?

they are more virulent, and therefore can be more toxic and important to identify.

what can we say about bacteria that have capsules?

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