1. Microbiologic Staining Method
顏 慕 庸
高 雄 榮 總

2. Microbiological Staining
Gram stain
Acid fast stain
KOH stain
Wet mount
Indian ink

3. Gram Stain Christian Gram, 1884 crystal violet iodine-KI alcohol wash
safranin

4. Significance
Culture
Buy time for golden period
Diagnosis

5. Interpretation Morphological Associate environment Site-specific
Immunity-oriented

6. Morphologic interpretation
cocci vs bacilli, cocco-bacilli
Gram positive vs Gram negative

7. Morphologic interpretation
cocci vs bacilli, cocco-bacilli
Gram positive vs Gram negative

8. Gram-Positive Cocci GPC in group GPC in chain GPC w/c capsule
GPC in pair
lancet shape
kidney shape
sarcinae

9. Gram-Positive Cocci GPC in group GPC in chain GPC w/c capsule
GPC in pair
lancet shape
kidney shape
sarcinae

10. Cocco-bacilli, Kidney-shape G (+) vs G (-)
Gram Negative Cocci
Cocco-bacilli, Kidney-shape
G (+) vs G (-)

11. Bacilli Gram positive bacilli spore spindle box car palisade beaded
branching
Gram negative bacilli
pleomorphic
cocco-bacilli
capsule
curved

12. Bacilli Gram positive bacilli spore spindle box car palisade beaded
branching
Gram negative bacilli
pleomorphic
cocco-bacilli
capsule
curved

13. Presumptive Diagnosis
Staphylococcus aureus
Streptococcus pneumoniae
Hemophilus influenzae
Klebsiella pneumoniae
Nocardia spp
Candida spp

14. 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

15. 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

16. 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

17. Site specific Gram negative cocci urine: CSF: sputum:
Gram positive bacilli
CSF:
sputum:
Gram positive cocci
urine:

18. Site specific Gram negative diplococci urine: N. gonorrheae
CSF: N. meningitidis
sputum: M. catarrhalis

19. Site specific Gram positive bacilli CSF: L. monocytogenes
sputum: Corynebaterium straitum
Gram positive cocci
urine: Enterococcus , Viridans strep.

20. Immunity - oriented Nosocomial infection Community-acquired 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

21. 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

22. Immunity - oriented T-cell immunity T-cell immunity GPB in CSF
L. monocytogenes
T-cell immunity
GNB in joint
Salmonella spp.

23. Acid Fast Stain Ziehl-Neelsen stain Kinyoun modification Mycobacteria
Nocardia, partial stain
NOT for actinomycosis