Microbiologic Staining Method

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Microbiologic Staining Method

• ? ? ?
• ? ? ? ?

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Microbiological Staining

• Gram stain
• Acid fast stain
• KOH stain
• Wet mount
• Indian ink

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Gram Stain

• Christian Gram, 1884
• crystal violet
• iodine-KI
• alcohol wash
• safranin

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Significance

• Culture
• Buy time for golden period
• Diagnosis

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Interpretation

• Morphological
• Associate environment
• Site-specific
• Immunity-oriented

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Morphologic interpretation

• cocci vs bacilli, cocco-bacilli
• Gram positive vs Gram negative

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Morphologic interpretation

• cocci vs bacilli, cocco-bacilli
• Gram positive vs Gram negative

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Gram-Positive Cocci

• GPC in group
• GPC in chain
• GPC w/c capsule

• GPC in pair
• lancet shape
• kidney shape
• sarcinae

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Gram-Positive Cocci

• GPC in group
• GPC in chain
• GPC w/c capsule

• GPC in pair
• lancet shape
• kidney shape
• sarcinae

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Gram Negative Cocci

• Cocco-bacilli, Kidney-shape
• G () vs G (-)

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Bacilli

• Gram positive bacilli
• spore
• spindle
• box car
• palisade
• beaded
• branching

• Gram negative bacilli
• pleomorphic
• cocco-bacilli
• capsule
• curved

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Bacilli

• Gram positive bacilli
• spore
• spindle
• box car
• palisade
• beaded
• branching

• Gram negative bacilli
• pleomorphic
• cocco-bacilli
• capsule
• curved

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Presumptive Diagnosis

• Staphylococcus aureus
• Streptococcus pneumoniae
• Hemophilus influenzae
• Klebsiella pneumoniae
• Nocardia spp
• Candida spp

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Associate environment

• Neutrophils, epithelial cells
• Aid for judgement of G(), G(-)
• Intracellular vs extracellular
• Mixed flora
• w/o PMN, w/o bacteria
• Non-bacteria organisms

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Associate environment

• Neutrophils, epithelial cells
• Aid for judgement of G(), G(-)
• Intracellular vs extracellular
• Mixed flora
• w/o PMN, w/o bacteria
• Non-bacteria organisms

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Associate environment

• Neutrophils, epithelial cells
• Aid for judgement of G(), G(-)
• Intracellular vs extracellular
• Mixed flora
• w/o PMN, w/o bacteria
• Non-bacteria organisms

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Site specific

• Gram negative cocci
• urine
• CSF
• sputum

• Gram positive bacilli
• CSF
• sputum
• Gram positive cocci
• urine

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Site specific

• Gram negative diplococci
• urine N. gonorrheae
• CSF N. meningitidis
• sputum M. catarrhalis

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Site specific

• Gram positive bacilli
• CSF L. monocytogenes
• sputum Corynebaterium straitum
• Gram positive cocci
• urine Enterococcus , Viridans strep.

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Immunity - oriented

• Nosocomial infection
• GNB in CSF
• cocco-bacilli in sputum
• T-cell immunity
• GPB in CSF

• Community-acquired
• GNB in CSF
• cocco-bacilli in sputum
• T-cell immunity
• GNB in joint

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Immunity - oriented

• Community-acquired
• GNB in CSF
• K. pneumoniae
• cocco-bacilli sputum
• H. influenzae

• Nosocomial infection
• GNB in CSF
• P. aeruginosa
• Enterobactericeae
• cocco-bacilli in sputum
• A. baumannii

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Immunity - oriented

• T-cell immunity
• GPB in CSF
• L. monocytogenes

• T-cell immunity
• GNB in joint
• Salmonella spp.

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Acid Fast Stain

• Ziehl-Neelsen stain
• Kinyoun modification
• Mycobacteria
• Nocardia, partial stain
• NOT for actinomycosis