microscopy - dl1.cuni.cz
TRANSCRIPT
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Microscopy
Tereza Kopecká
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Remember!
• Microbiologists love microscopy.
• We perform it always, if possible and informative.
• If it‘s possible to identify through microscopyalone, we take it. (typically parasitology and mycology)
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What do we need for microscopy?
• A microscope – let‘s focus on the light one.
• Slides– Native (with saline solution or KOH)
– stained• Gram stain – bacteria, yeasts
• Ziehl-Neelsena – mycobacteria, nocardiae, cryptosporidia
• Giemsa – pneumocysts, parasites
• Lactophenol blue - molds
• India ink (Burri stain) – capsules – Cryptococcus
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What do we observe?
• New samples
• Cultures
• Positive blood cultures
– Blood sample from a septic patient
– Blood culture bottle
– Incubation and detection of chemical changesinside
– BD Bactec or BactALERT signalizes positive
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New samples
• The liquid ones. The only swab is the MOP
– Sputum and other respiratory
– CSF
– Pus, liquid contents of cavities, etc.
• Bacteriologists never look at stools but
– Parasitologists do (+tape!)
– Virologists as well (electron microscope)
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Why observe cultures?
• It might help with identification
– If we have MALDI-TOF, no need
– Not every lab has a MALDI, for chemicalidentification, basic morphology is needed
• In molds, it‘s absolutely necessary
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Why observe positive blood cultures?
• We reassure ourselves about real positivity– Do you see no bacteria? Something is wrong
– Let‘s check the curves at Bactec
• We obtain basic data about the microbes– Immediate treatment
– An impulse for other identification methods• Sprout hyphae in yeasts – 2 hours
– A correct susceptibility testing• MH agar with horse blood in cocci forming chains
• Correct sets of antibiotics
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Sputum microscopy
• We are interested in:– Human cells
– microbes
• Gram stain is the basic one
• Human cells show us:– Sample validity (Bartlett score)
• With more leukocytes and less squamous cells, thesputum is better
– Intensity of the inflammation• The appearance is quite different in abscess and mild
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Sputum microscopy
• We can see the agents!
– Lancet-like G+ diplococci
– G- cocci in tetraplets
– Tiny G- coccobacilli
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Sputum microscopy for TB
• Can be informative but must be targeted
– Different staining method (Ziehl-Neelsen)
– Lower sensitivity
• Negative slide does not mean a healthy patient
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CSF microscopy
• Routine or suspect neuroinfection
• Quantity of leukocytes and the appearance ofthe microbes must be reported to thephysician
• If they are not present, it‘s an impulse forother methods like PCR or serology
• CSF microscopy might be a life-savingprocedure
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A special topic: sprout hyphae
• Yeasts put in horse serum for 2 hours
• If it‘s Candida albicans, broad hyphae are formed
• Mycology practicals…
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Let‘s go!