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Microorganism
Staining
| Question | Answer |
|---|---|
| Cocci bacteria arrangements: | diplococci - paired, staphylococci - grape-clusters, streptococci - chains |
| Some pathogenic cocci include: | Staphylococcus aureus (TSS), streptococcus pneumonia, Neisseria gonorrhea, Neisseria meningitidis, methicillin-resistant staph aureus |
| Bacilli bacteria | Rod-shaped bacteria; pathogenic: Clostridium tetani, C. botulinum, C. difficile, C. perfringens (food poisoning), bacillus anthracis |
| Coccobacilli bacteria | Rod-shaped, short & wide bacteria; pathogenic: Hemophilus influenza, Chlamydia trachomatis |
| Gram-negative bacteria: | Don't stain; Salmonella, Shigella dysenteriae, E. coli, Pseudomonas aeruginosa (pneumonia, UTI), N. meningitidis, N. gonorroeae, S. aureus |
| Gram-positive bacteria: | Stain blue due to cell wall of teichoic acid, include - Clostridium botulinum, C. tetani, Corynebacterium diptheriae, Actinomyces |
| Spirochetes are: | Spiral/corkscrew bacteria; pathogenic: Leptospira (Weil's disease), Triponema pallidium (syphilis), Borrelia burgdorferi (Lyme disease), argyrophil stains |
| Some bacteria types that don't have typical bacterial attributes include: | Rickettsiae, mycoplasmas |
| Which bacteria can only reproduce w/i living host cell? | Rickettsiae & chlamydiae |
| Rickettsiae causes: | Rocky mountain spotted fever, typhus, tick-borne diseases |
| Mycoplasmas: | Don't form cell walls, can cause pneumonia, still birth, abortion |
| Which stains are commonly used to screen for the presence of bacteria? | Methylene blue, Giemsa |
| Mycosis is: | disease caused by fungi |
| Mycology is: | study of fungi |
| Molds are: | Filamentous fungi, easy to see septate hyphae in Aspergillus fumigatus |
| Yeasts are: | Single round/oval "budding" cells, Cryptococcus neoformans (meningitis) |
| Yeast-like fungi are: | Filamentous, reproduce by budding to form pseudohyphae, Candida albicans (yeast infection, thrush) |
| Dimorphic fungi: | Have yeast-like morphology at 37 °C, filamentous morphology on artificial media @ 25°C, Blastomyces dermatitidis, Coccidioides immitis (Valley fever), Histoplasma capsulatum |
| Viruses are: | Protein-coated RNA or DNA, seen w/ EM, may form cellular inclusions visible w/ light microscope, e.g. Negri bodies of rabies & cytomegalovirus |
| Protozoans are: | Single-celled microorganisms, psuedopods, flagella, or cilia, Entamoeba histolytica, Giardia lamblia, Toxoplasma gondii |
| Pneumocystis carinii: | aka Pneumocystis jirovecii, used to be considered protozoan, more closely related to yeast |
| Acid-fast bacteria are: | Mycobacteria, rod-shaped, may exhibit filamentous growth, cell walls have lots of lipids so once stained resists acid decolorization, Mycobacterium tuberculosis, mycobacterium leprae |
| Kinyoun & Ziehl-Neelsen stains facts | Detect presence of acid-fast mycobacteria, 10% NBF, no Carnoy, QC: tissue w/ acid-fast organisms |
| Kinyoun stain solutions | Kinyoun Carbol-Fuchsin + acid alcohol + methylene blue |
| Kinyoun & Ziehl-Neelsen stain results | acid-fast bacteria: bright red - background: light blue |
| When cutting slides to test for acid-fast bacteria must use: | Millipore filtered water in bath, no tap or dH2O, use negative control from same day/bath/microtome |
| Kinyoun Carbol-Fuchsin solution | basic fuchsin + phenol crystals (enhance stain) + 95% + filtered water filter before use |
| Methylene blue solution | methylene blue + 95% + H2O |
| Kinyoun vs. Ziehl-Neelsen Carbol-Fuchsin solutions | Kinyoun has more basic fuchsin & phenol than Ziehl-Neelsen |
| Ziehl-Neelsen method solutions | Ziehl-Neelsen Carbol-Fuchsin + acid alcohol + methylene blue |
| Ziehl-Neelsen Carbol-Fuchsin solution | basic fuchsin + phenol crystals + 100% + filtered H2O filter before use |
| Microwave Ziehl-Neelsen method solutions | Carbol-Fuchsin soln. + acid alcohol + methylene blue w/ acetic acid |
| Microwave Ziehl-Neelsen method results | acid-fast bacteria (incl. mycobacterium avium intracellulare): red - rbc: pink - mast cells: blue - other: pale blue |
| Carbol-Fuchsin solution | pararosaniline (basic fuchsin) + phenol crystals + isopropanol + filtered H2O filter before use |
| Fite stain facts | To detect presence of Mycobacterium leprae, 10% NBF, no Carnoy, QC: section w/ leprosy organisms |
| Fite stain solutions | xylene-peanut oil + acid alcohol + Ziehl-Neelsen Carbol-Fuchsin soln. + methylene blue w/ acetic acid |
| Fite stain results | M. leprae, acid-fast bacteria: bright red - background: light blue |
| Xylene-peanut oil is used to: | Protect capsule of leprae organism from dissolving during deparaffinization |
| Microwave auramine-rhodamine fluorescence technique facts | To detect presence of Mycobacterium tuberculosis & acid-fast bacteria, 10% NBF QC: section w/ acid-fast mycobacteria |
| Microwave auramine-rhodamine fluorescence technique solutions | auramine O-rhodamine B soln. + acid alcohol (decolorize) + Erichrome black T 0.3% (quench autofluorescence) |
| Microwave auramine-rhodamine fluorescence technique results | acid-fast bacteria: reddish-yellow fluorescence - background:black |
| Auramine O-rhodamine B solution | auramine O (fluorescent dye) + rhodamine B (fluorescent dye) + glycerol + phenol + filtered H2O |
| Alcian yellow-T blue facts | To ID H. pylori, no glyoxal QC: tissue w/ H. pylori |
| Alcian yellow-T blue solutions | periodic acid (ox. cell wall) + alcian yellow (stain mucin) + T blue (stain wall) + sodium metabisulfite |
| Alcian yellow-T blue results | H. pylori: deep blue - mucin: yellow |
| Gram stains purpose | ID Gram+ & - bacteria, QC: section w/ Gram +& - bacteria |
| Gram stains solutions | crystal violet (stain all bacteria) + basic fuchsin (stain Gram -) + Gram iodine (mordant) + 100% or acetone (diff.) + tartrazine or picric acid-acetone soln. (counterstain) if overdecolorize, Gram + no stain |
| Gram stains results | Gram + : purple - Gram - : red |
| Brown-Hopps modification of Gram stain solutions | crystal violet in dH2O + basic fuchsin + Gram iodine + Gallego solution + picric acid-acetone |
| Brown-Hopps modification of Gram stain results | Gram + : blue - Gram - : red - background: yellow - nuclei: light red |
| Gallego solution | formalin + glacial acetic acid + dH2O |
| Giemsa stains can demo: | Bacteria, rickettsias, Toxoplasma gondii |
| Modified Diff-Quik Giemsa Stain purpose | ID H. pylori, QC: section w/ H. pylori, Romanowsky-type |
| Modified Diff-Quik Giemsa Stain solutions | Diff-Quik soln. I + Diff-Quik soln. II + acetic acid water |
| Modified Diff-Quik Giemsa Stain results | H. pylori, nuclei: dark blue - other bacteria: blue - cytoplasm: pink |
| Hotchkiss-McManus PAS reaction, Gridley, & Grocott-methenamine-silver nitrate stains purpose | Demo fungi, QC: section w/ fungi |
| Hotchkiss-McManus PAS reaction solutions | periodic acid + Schiff reagent + sulfurous rinse (remove excess leucofuchsin) + fast green w/ acetic acid (or light green) |
| Hotchkiss-McManus PAS reaction results | fungi: rose - background: green |
| Gridley stain solutions | chromic acid + Schiff + aldehyde fuchsin (reacts w/ Schiff like an aldehyde) + metanil yellow |
| Gridley stain results | Mycelia, elastic fibers, mucin: deep purple - conidia: deep rose to purple - background: yellow |
| Grocott-methenamine-silver nitrate stain facts | 4-5 μm paraffin, 6 μm frozen, QC: for Pneumocystis use Pneumocystis control, use chemically clean glassware & nonmetallic forceps |
| Grocott-methenamine-silver nitrate stain solutions | chromic or periodic acid (ox. walls to aldehyes) + methenamine-silver nitrate working + gold chloride + sodium bisulfite (remove acid) + sodium thiosulfate + light green w/ acetic acid |
| Grocott-methenamine-silver nitrate stain results | fungi: sharply delineated in black - mucin: taupe to dark gray (if black, methenamine too hot) - background: green |
| Microwave methenamine-silver nitrate procedure purpose | Best to demo on cytospin preps or frozen diagnosis Pneumocystis carinii |
| Microwave methenamine-silver nitrate procedure facts | cytospin prep fixed in 95%, frozen fixed in 40% formaldehyde, 6 μm, paraffin routine QC: pneumatocyst for P. carinii, fungi for fungi |
| Microwave methenamine-silver nitrate procedure solutions | chromic acid + methenamine-silver nitrate working + gold chloride + sodium thiosulfate + sodium bisulfate (clear) + light green w/ acetic acid |
| Microwave methenamine-silver nitrate procedure results | fungi & P. carinii: black - background: green |
| Methenamine-silver working solution | methenamine + AgNO3 + Borax + dH2O |
| What stains can demo Crytococcus neoformans? | Mayer mucicarmine & alcian blue; may demo Blastomyces dermatitidis & Rhinosporidium seeberi |
| Warthin-Starry, microwave Warthin-Starry, & Dieterle purpose | Demo spirochetes, legionella, QC: section w/ spirochetes or legionella |
| Warthin-Starry Technique solutions | AgNO3 2% w/ acidulated water + AgNO3 w/ acidulated water, gelatin soln. & hydroquinone |
| Warthin-Starry Technique results | spirochetes, other bacteria: black - background: pale yellow to light brown |
| Acidulated water | triple-distilled H2O + citric acid pH4.0 |
| Microwave Warthin-Starry Method solutions | glycine-acetic acid pH 4.2 + silver nitrate-gelatin-hydroquinone developer w/ glycine-acetic acid |
| Microwave Warthin-Starry Method results | alipia felis (cat-scratch bacillus), legionella pneumophila, nocardia asteroides, H. pylori: black - nuclei, rbc: brown |
| Dieterle Method solutions | alcoholic uranyl nitrate + alcoholic gum mastic (protect organism) + AgNo3 + developer + formic acid |
| Dieterle Method results | spirochetes, bacteria: brown to black - background: pale yellow or tan Melanin, chromatin, formalin pigment, foreign materials may also stain brown to black |
| Dieterle developer solution | alcoholic gum mastic + acetone + hydroquinone + pyridine + sodium sulfite + formaldehyde + dH2O |
| Microwave Steiner & Steiner procedure purpose | Demo spirochetes, H. pylori, legionella QC: section w/ what staining for |
| Microwave Steiner & Steiner procedure solutions | uranyl nitrate (sens.) + AgNo3 + gum mastic + reducing soln. |
| Microwave Steiner & Steiner procedure results | Spirochetes, H. pylori, Legionella, other nonfilamentous bacteria - dark brown to black background - light yellow |
| Microwave Steiner & Steiner reducing solution | gum mastic + hydroquinone (red.) + abs. alcohol |
| When staining for spirochetes avoid what? | Mercurial & chromate fixatives, 10% NBF preferred |
| Sulfurous rinse solution | HCl + potassium metabisulfite + dH2O |
| Argentaffin stains | Gomori methenamine-silver, Grocott methenamine-silver, Fontana Masson, von Kossa |
| Argyrophil stains | Reticulin, Holmes, Bielschowsky, Warthin-Starry, Dieterle, Steiner & Steiner, Grimelius, Churukian-Schenk |
| Sayeed's Stain purpose & reagents | Detect H. Pylori, periodic acid + Schiff + Mayer's hemalum |
| Plasmodium parasites | Cause malaria which deposits pigment closely related to formalin pigment |
| Cryptosporidium | Protozoa, causes severe outbreaks of diarrhea among AIDS patients |
| Listeria monocytogenes | Cause rare form of meningitis, stain variably with Gram stain, appear as intracellular rods in "Chinese letter" formation |
| Ziehl-Neelson may also demonstrate | Lipofuscins if technique is modified to include heat and prolonged staining |
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