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Microorganisms
Microorganisms in the histology lab
| Question | Answer |
|---|---|
| Bacteria | Tiny, single celled organisms |
| Bacteria | ~0.2-10 micrometers |
| Spherical or ovoid bacteria | Cocci |
| Cause of TSS (Toxic Shock Syndrome) | Staph Aureus |
| Cocci | Staph aureus, Streptococcus pneumoniae, Nisseria gonorrhoeae, and Neisseria meningitdis |
| Rod shaped bacteria | Bacilli |
| Bacilli | Clostridium tetani, Clostridium botulinum, and bacillus anthracis. |
| Coccobacilli | Haemophilus influenzae, and Chlamydia trachomatis |
| Spiral shaped bacteria | Spirochetes |
| Spirochetes | Syphillus:Treponema pallidum, and Borrelia burgdorferi: Lyme disease |
| What stains are primarily used to identify Spirochetes? | Silver stains |
| Stain for presence of bacteria | Giemsa and Methylene Blue |
| Acid fast techniques are of value in the detection of what? | Mycobacteria, rod shaped organisms that sometimes exhibit filamentous (fungus like) growth |
| Fungi | Unicellular or multicellular primitive plants that have a distinct membrane bound nucleus containing genetic material. |
| Filamentous fungi are also called | molds |
| Yeasts | Single round or oval cells that reproduce by "budding" |
| Classified as a yeast | Crytococcus neoformans |
| Yeastlike fungi that does not detach from parent cell "pseudohyphae" | Candida albicans |
| Virus | Composed of either DNA or RNA protein |
| Virus | Protein coated genes that need living cells to provide energy and the machinery for duplication. |
| Virus | ~20-30 nm |
| Most viruses can only be seen with a _ microscope. | Electron |
| Protozoans | Single celled microorganisms that are functionally complex structures |
| Protozoans | Entamoeba histolytica, Giardia lamblia, and Toxoplasma gondii. |
| Kinyoun Acid Fast | Detect presence of Acid-fast bacteria |
| Kinyoun Acid Fast | Lipoid capsule or organism takes up carbol fuchsin and deists decolorization with dilute mineral acid.Acid alcohol used to decolorize to get an even decolorization. |
| Kinyoun Acid Fast | 10% NBF (Carnoy can't be used) |
| Kinyoun Acid Fast | 4-5 micrometers |
| Kinyoun Acid Fast control | Tissue containing acid fast organisms |
| Kinyoun Acid Fast | 1. Carbol fuchsin 2. Water 3. 1% acid alcohol 4. water 5. Methylene Blue 6. rinse 7. Dehydrate and clear |
| Kinyoun Acid Fast results | Acid-fast bacteria: Bright red Background: Blue |
| Ziel-Neelsen | Detection of acid fast bacteria |
| Ziel-Neelsen | Afixed tissue with exception of Carnoy |
| Ziel-Neelsen | 4-5 micrometers |
| Ziel-Neelsen control | Tissue containing acid fast organisms |
| Ziel-Neelsen results | Acid-fast bacteria: Bright red Background: Blue |
| Ziel-Neelsen microwave results | Acid fast organisms: Red Erythrocytes: Pink Mast cells: Blue Other tissue elements: Pale blue |
| Fite | Detection of mycobacterium leprae |
| Fite | Lipoid capsule of organism takes up carbol fuchsin and resists decolorization with dilute mineral acid. |
| Fite | 10% NBF and others with exception of Carnoy |
| Fite | 4-5 micrometers |
| Fite control | Tissue containing leprosy organisms |
| Fite | 1. Xylene peanut oil 2. Blot 3. Ziel Nelson carbol fuchsin 4. wash 5. 1% acid alcohol 6. Wash 7. Methylene blue 8. Rinse 9. Blot 10. Mount with synthetic resin |
| Fite results | M leprae and other acid fast bacteria: Bright red Background light blue |
| Auramine Rhodamine | Detection of M tuberculosis or other acid fast organisms |
| Auramine Rhodamine | 10% NBF |
| Auramine Rhodamine | 4-5 micrometers |
| Auramine Rhodamine control | Tissue containing Acid fast mycobacteria |
| Auramine Rhodamine | 1. Auramine rhodamine 2. rinse 3. Acid alcohol 4. rinse 5. 0.3% eriochrome black T 6. Rinse 7. Dry 8. Dip in xylene and mount |
| Auramine Rhodamine results | Acid fast organisms: Reddish-yellow fluorescence Background: Black |
| Gram stain (Brown-Hopps) | Demonstration of gram positive and gram negative organisms |
| Gram stain (Brown-Hopps) | 10% NBF |
| Gram stain (Brown-Hopps) | 4-5 micrometers |
| Gram stain (Brown-Hopps) control | Sections containing bothe Gram positive and gram negative organisms should be used |
| Gram stain (Brown-Hopps) | 1. Crystal violet 2. rinse 3. iodine 4. rinse 5. decolorize 6. rinse 7. basic fuchsin 8. rinse 9. Gallego solution 10. rinse 11. Acetone 12. Picric acid-acetone 13. Acetone 14. Acetone-xylene 15 Mount |
| Gram stain (Brown-Hopps) results | Gram +: blue-black Gram -: Red Background tissue: Yellow Nuclei: light red |
| Giemsa can be used to demonstrate | T gondii, rickettsias, and bacteria |
| Diff Quick | Identification of H pylori |
| Diff Quick | Romanowsky stains "neutral" dyes combing basic and acid dyes creates a wide range of color. |
| Diff Quick | 10% NBF |
| Diff Quick | 4-5 micrometers |
| Diff Quick control | Tissue with H pyori |
| Diff Quick | 1. Diff Quick solution I 2. Diff Quick solution II 3. Rinse 4. Acetic water 5. Rinse 6. Dehydrate, clear, and mount |
| Diff Quick results | H pylori: Dark blue Other bacteria: Blue Nuclei: Dark Blue Cytoplasm: Pink |
| Solution I in Diff Quick | Buffered solution of Eosin Y (Anionic Dye, stains cytoplasmic elements pink) |
| Solution II in DIff Quick | Cationic dye mixture of azure A and methylene blue, stains nuclei and bacteria blue |
| Alcian yellow-toulidine blue | Detection of H pylori |
| Alcian yellow-toulidine blue | Alcian yellow is a monoazo dye that reacts similar to alcian blue, staining mucin yellow. Toulidine blue is a basic dye and metachromatic stain that stains H pylori and nuclei blue |
| Alcian yellow-toulidine blue | 10% NBF |
| Alcian yellow-toulidine blue | 4-5 micrometers |
| Alcian yellow-toulidine blue control | Tissue containing H pylori |
| Alcian yellow-toulidine blue | 1. Periodic acid 2. wash 3. Sodium metabilsulfite 4. rinse 5, Alcian yellow 6. wash 7. Toluidine blue 8. wash 9. blot 10 dehydrate, clear, mount |
| Alcian yellow-toulidine blue results | H pylori: Blue Mucin: Yellow Background: Pale blue |
| Hotchkiss-McManus periodic acid Schiff | Demonstration of fungi |
| Hotchkiss-McManus periodic acid Schiff | Polysaccharides present in the fungal cell walls are oxidized by the periodic acid to aldehydes. Aldehydes react with Schiff reagent to yield rose colored fungi. |
| Hotchkiss-McManus periodic acid Schiff | 10% NBF, Bouin or Zenker |
| Hotchkiss-McManus periodic acid Schiff | 4-5 micrometers |
| Hotchkiss-McManus periodic acid Schiff control | Section containing fungi |
| Hotchkiss-McManus periodic acid Schiff | 1. periodic acid 2. rinse 3. Schiff 4. rinse with sulfurous acid 5. wash 6. counterstain light green 7. rinse 8. Dehydrate, clear and mount |
| Chromic acid Schiff | Demonstration of fungi |
| Chromic acid Schiff | 10% NBF |
| Chromic acid Schiff | 4-5 micrometers |
| Chromic acid Schiff control | Section containing fungi |
| Chromic acid Schiff | 1., Chromic acid 2. rinse 3. Schiff 4. rinse in sulfurous acid 5. rinse 6. Counterstain with harris heme or light green 7. wash 8. Dehydrate, clear and mount |
| Chromic acid Schiff results | Fungi: Deep rose to purple Nuclei if heme is used: Blue Background in light green is used: Green |
| Gridley | Demonstration of fungi |
| Gridley | 10% NBF |
| Gridley | 4-5 micrometers |
| Gridley control | Section containing fungi |
| Gridley | 1. Chromic Acid 2. rinse 3. schiff 4. rinse 5. 70% alcohol 6. Aldehyde fucchsin 7. rinse in 95% alcohol 8. rinse 9. counterstain with mental yellow 10. rinse 11. Dehyrate, clear, mount |
| Gridley results | Mycelia: Deep purple Conidia: Deep rose to purple Background: Yellow Elastic fibers: Deep purple |
| Grocott Methenamine Silver Nitrate | Demonstration of fungi |
| Grocott Methenamine Silver Nitrate | 10% NBF |
| Grocott Methenamine Silver Nitrate | 4-5 micrometers |
| Grocott Methenamine Silver Nitrate control | Section containing fungi |
| Grocott Methenamine Silver Nitrate | 1. Chromic Acid 2. rinse 3. Sodium bisulfite 4. rinse 5. Methnamine silver 6. rinse 7. gold chloride 8. rinse 9. sodium thiosulfate 10 rinse 11. counterstain 12. dehydrate clear mount |
| Grocott Methenamine Silver Nitrate results | Fungi: Crisp black Mucin: Taupe to dark grey Background green |
| Mayer mucicarmine & Alcian Blue stains for | C neoformans, Blastomyces dematidis, and Rhinosporidium |
| Warthin-Starry | Demonstration of spirochetes |
| Warthin-Starry | Argyrophil method Hydroquinone is used to reduce |
| Argyophil | Have the ability to bind silver ions from a solution but they do not have the ability to reduce the silver to a visible form. |
| Argentaffin | Have the ability to bind and reduce silver ions from a solution |
| Warthin-Starry | 10% NBF |
| Warthin-Starry | 4-5 micrometers |
| Warthin-Starry control | Tissue that contains spirochetes |
| Warthin-Starry | 1. Silver Nitrate 2. Developer 3. wash 4. Dehydrate clear mount |
| Warthin-Starry results | Spirochetes: Black Other bacteria: black Background: Pale yellow to light brown |
| If sections are over developed they can be treated with what and then restrained? | Iodine or sodium thiosulfate for color removal |
| Dieterle | Demonstration of Spirochetes and Legionella organisms |
| Dieterle | Argyrophillic |
| Dieterle | 10% NBF |
| Dieterle | 4-5 micrometers |
| Dieterle control | Tissue containing spirochetes or legionella |
| Dieterle | 1. Alcohol uranyl nitrate 2. rinse 3. 95% 4. Distilled Water 5. Silver nitrate 6. rinse 7. Developer 8. rinse 9. 10% formic acid 10. rinse 11. dehydrate, clear, mount |
| Dieterle results | Spirochetes, bacteria: Brown to black Background: pale yellow or tan |
| Steiner & Steiner | Demonstation of spirochetes, h pylori, or legionella |
| Steiner & Steiner | Argyrophillic |
| Steiner & Steiner | 10% NBF (Chromate fixatives should be avoided) |
| Steiner & Steiner | 4-5 micrometers |
| Steiner & Steiner control | Tissue with spirochetes, H pylori, or L pnuemonphila must be used |
| Steiner & Steiner | 1. Uranyl nitrate 2, rinse 3. silver nitrate 4. rinse 5. 95% 6. 100% 7. Gum mastic 8. Air dry 9. rinse 10. reduce 11. rinse 12. dehydrate clear mount |
| Steiner & Steiner results | Spirochetes, H pylori, L pnuemophila and other non filamentous bacteria: Dark brown to black Background: Light yellow |
Created by:
Ziek98
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