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43 terms

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?