{"id":901,"date":"2018-08-04T04:58:42","date_gmt":"2018-08-04T04:58:42","guid":{"rendered":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/?p=901"},"modified":"2018-08-13T21:35:42","modified_gmt":"2018-08-13T21:35:42","slug":"microbiology-lab-procedure-notes","status":"publish","type":"post","link":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/microiology\/microbiology-lab-procedure-notes\/","title":{"rendered":"Microbiology Lab Procedure Notes"},"content":{"rendered":"

Microbiology Lab Procedure Notes By \u00a92018 Shirley S. Chung, Green River Community College<\/strong><\/p>\n

Download these notes.<\/a><\/p>\n

Smear<\/strong><\/span><\/h2>\n
    \n
  1. Clean slide<\/li>\n
  2. Label: my name, organism, date, medium<\/li>\n<\/ol>\n

    Broth Smear<\/u><\/strong><\/h2>\n
      \n
    1. Agitate broth unless otherwise stated<\/li>\n
    2. FLAME, 1 loopful in center of slide spread to dime-size.<\/li>\n
    3. FLAME, 2nd loopful in center of slide spread to dime-size.<\/li>\n
    4. FLAME<\/li>\n<\/ol>\n

      Solid Smear<\/u><\/strong><\/h2>\n
        \n
      1. FLAME, 1 loopful water (or drop)<\/li>\n
      2. FLAME, 2nd loopful water<\/li>\n
      3. FLAME 1 loopful specimen and mix well in water.<\/li>\n
      4. FLAME 2nd loopful specimen if needed and mix well in water.<\/li>\n
      5. FLAME<\/li>\n
      6. Air Dry<\/li>\n
      7. Heat Fix<\/strong><\/li>\n
      8. Use clothespin.<\/li>\n
      9. Pass slide’s underside over the flame (coolest part of flame)<\/li>\n
      10. 3 passes<\/li>\n<\/ol>\n

        Simple Stain<\/span> (Methylene blue, Crystal Violet, Carbol Fuchsin)<\/strong><\/h2>\n
          \n
        1. Smear and heat fix.<\/li>\n
        2. Flood stain<\/li>\n<\/ol>\n

          Carbol Fuchsin, 15-30 sec<\/p>\n

          Crystal Violet, 20-60 sec<\/p>\n

          Meth. Blue, 1-2 min<\/p>\n

            \n
          1. Rinse with Di Water<\/li>\n
          2. Bibulous blot, Do not wipe<\/li>\n<\/ol>\n

            Gram Stain<\/strong><\/span><\/h2>\n
              \n
            1. Flood with Crystal Violet<\/li>\n
            2. Wait 1 min<\/strong><\/li>\n
            3. Flush with Di water<\/li>\n
            4. Flood with Gram’s Iodine mordant<\/li>\n
            5. Wait 1 min<\/strong><\/li>\n
            6. Flush with Di water<\/li>\n
            7. Decolorize<\/li>\n
            8. Flush immediately<\/strong><\/li>\n
            9. Counterstain Safranin<\/li>\n
            10. Wait 45 sec<\/strong><\/li>\n
            11. Flush with Di water<\/li>\n
            12. Bibulous Blot<\/li>\n<\/ol>\n

              Acid-Fast Staining<\/strong><\/span><\/h2>\n
                \n
              1. Smear and heat fix.<\/li>\n
              2. Flood smears with carbol fuchsin<\/li>\n
              3. Place over a beaker of water on a warm hot plate<\/li>\n
              4. Steam 5 minutes<\/strong> (do NOT allow stain to evaporate, replenish as needed)<\/li>\n<\/ol>\n

                OR<\/u><\/strong><\/p>\n

                Flood smear with carbol fuchsin WITH TERGITOL<\/u> for 5-10 minutes<\/strong><\/p>\n

                  \n
                1. If slides are heated, allow to cool.<\/li>\n
                2. Flush with Di water<\/li>\n
                3. Decolorize with acid-alcohol<\/li>\n
                4. Flush with Di water<\/li>\n
                5. Counterstain with Meth. Blue for 2 min<\/strong><\/li>\n
                6. Flush with Di water<\/li>\n
                7. Bibulous blot.<\/li>\n<\/ol>\n

                  Spore Staining<\/strong><\/span><\/h2>\n
                    \n
                  1. Smear and heat fix<\/li>\n
                  2. Flood with malachite green<\/strong><\/li>\n
                  3. Place on beaker of water sitting on hot plate<\/li>\n
                  4. Steam for 2-3 minutes<\/strong> (do NOT allow stain to evaporate, replenish as needed)<\/li>\n
                  5. Remove from hot plate<\/li>\n
                  6. Cool<\/li>\n
                  7. Flush with Di water (this is the decolorizing step)<\/li>\n
                  8. Counterstain Safranin for 30 seconds<\/strong><\/li>\n
                  9. Flush with Di water<\/li>\n
                  10. 10. Bibulous blot<\/li>\n<\/ol>\n

                     <\/p>\n

                    Microbiology Lab Procedure Notes By \u00a92018 Shirley S. Chung, Green River Community College<\/strong><\/p>\n","protected":false},"excerpt":{"rendered":"

                    Microbiology Lab Procedure Notes By \u00a92018 Shirley S. Chung, Green River Community College Download these notes. Smear Clean slide Label: my name, organism, date, medium Broth Smear Agitate broth unless otherwise stated FLAME, 1 loopful in center of slide spread to dime-size. FLAME, 2nd loopful in center of slide spread to dime-size. FLAME Solid Smear […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[36],"tags":[],"class_list":["post-901","post","type-post","status-publish","format-standard","hentry","category-microiology"],"_links":{"self":[{"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/posts\/901","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/comments?post=901"}],"version-history":[{"count":6,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/posts\/901\/revisions"}],"predecessor-version":[{"id":1110,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/posts\/901\/revisions\/1110"}],"wp:attachment":[{"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/media?parent=901"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/categories?post=901"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/integrativewellnessandmovement.com\/iwmbasicscience\/wp-json\/wp\/v2\/tags?post=901"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}