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tissues
performed using touch preparations
swabs
must be rolled back and forth across a dry, clean slide
single-drop smear
turbid
high numbers of organisms are expected
specimen is thick
centrifuged smear
used when a small number of organisms are expected
often clear
layered smear
used for low-volume specimens
a drop is placed on a slide and allowed to dry, then another drop is placed on top of the dried smear
albumin
may be added to CSF to help cellular material adhere to the slide
mucolytic agents
may be added to extremely mucoid specimens
gram stain
places bacteria into one of the two groups (g+/g-)
crystal violet
primary stain, gram staining
Gram’s iodine
mordant, gram staining
Acetone-alcohol
decolorizer, gram staining
Safranin red
counterstain, gram staining
acid-fast staining
used primarily to detect Mycobacterium species
kinyoun and ziehl-neelsen
two methods of Fuchsin stain
kinyoun method
also known as the cold method; uses a higher concentration of detergent to facilitate staining
Ziehl-Neelsen method
hot method; uses heat to facilitate staining and recommended for pulmonary TB
carbolfuchsin
primary stain, fuchsin staining
acid-alcohol
decolorizer, fuchsin staining
methylene blue
counterstain, fuchsin staining
malachite green
alternative for methylene blue
red
positive color for acid-fast staining
negative color for acid-fast staining
blue
auramine-rhodamine
acid-fast organisms fluoresce yellow or orange under a fluorescent microscope
methylene blue
used to observe metachromatic granules in Corynebacterium diphtheriae
acridine orange
fluochrome dye that stains both g+ and g- organisms both living and dead; used for low bacterial yield
saline mount
used to determine the biologic activity of microorganisms
hanging-drop procedure
same purpose as a saline mount but with a deeper field of focus and without the distorting effects from the coverslip’s weight; used to observe motility
darkfield examination
used to visualize organisms that are invisible by brightfield microscopy with great difficulty; demonstrates spirochetes
0
+1
+2
+1
give me the grades for the following (neutrophil edition)
<10
10-25
>25
presence of mucus
-1
-2
give me the grades for the following (eosinophil edition):
10-25
>25
liquid
semi-solid
solid
three kinds of culture media based on consistency/physical state
0.5%-1.0%
How much agar is in a semi-solid culture media?
2%-3% agar
amount of agar in solid culture media
agar
polysaccharide extract of seaweed/algae
synthetic
non-synthetic
cell culture
three kinds of culture media based on composition
plated media
dispensed in sterile petri dishes
tubed media
dispensed in sterile test tubes
non-selective media
supports most non-fastidious bacteria
selective media
support the growth of one type of bacteria and not the other; may contain inhibitory substances
differential media
allows the grouping of bacteria on the basis of different characteristics displayed on the medium; may be selective or non-selective
enriched media
growth enhancers have been added
enrichment broth (selective enrichment)
designed to encourage the growth of small numbers of a particular organism while suppressing the other flora present in the specimen
transport media
prevents replication while keeping bacterial cells viable
blood agar plate
most commonly used media
for the cultivation of fastidious bacteria and demonstration of hemolytic reactions
chocolate agar
dark brown in color
blood agar except the RBCs are already lysed
for the cultivation of Haemophilus and Neisseria
Thayer-Martin agar
modified chocolate agar
selective for Neisseria gonorrhoeae and Neisseria meningitidis
antibiotics are added
can be modified to have trimethoprim
Proteus spp.
What does trimethoprim inhibit?
Martin Lewis Agar
has vancomycin, colistin, and anisomycin (to inhibit yeast)
eosin methylene blue agar
used for the isolation and differentiation of lactose fermenting and non-lactose fermenting enteric bacilli
inhibits g+ bacteria
Mac Conkey agar
same use as EMB
contains bile salts and crystal violet to inhibit g+ bacteria
neutral red functions as an inhibitor
Mannitol Salt agar
for the selective isolation of coagulase positive staphylococci
7.5% NaCl inhibits most bacteria
phenol red is the indicator used
phenyl ethyl alcohol agar
selective for the isolation of gram+ cocci
inhibits g- bacteria
has blood
selenite broth (Sel-F)
selective for Salmonella and some strains of Shigella
should not be autoclaved
toxic for most Enterobacteriaceae
thioglycolate broth
supports the growth of aerobes, microaerophiles, and anaerobes
contains Resazurin which functions as an oxidation-reduction indicator
no more than twice autoclaved
if green color has spread it must not be used
xylose lysine desoxycholate (XLD) agar
for isolation and differentiation of Salmonella and Shigella from other g- enteric bacteria
should not be autoclaved
high NaCl content inhibits most enterobacteriae and g+ bacteria
contains ferric ammonium citrate to visualize H2S production
phenol red as pH indicator
hektoen enteric agar
selective and differential of Salmonella and Shigella
similar utility as that of XLD
brain heart infusion
supportive medium for a wide variety of organisms
general purpose medium
lowenstein-jensen medium
selective enrichment for Mycobacterium spp.
malachite green inhibits growth of other bacteria
contains homogenized eggs for production of niacin
mueller-hinton
used for antimicrobial susceptibility testing
chromagar
provides a variety of colors for differentiation of specific isolates
streaking
pour plate
spread plate
three kinds of inoculation techniques for isolation