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This Concept Map, created with IHMC CmapTools, has information related to: LAB 5 - DIRECT STAIN AND INDIRECT STAIN, Place the slide on a staining tray and cover the entire film with crystal violet. Stain for 30 seconds. step 4 Pick up the slide by one end and hold it at an angle over the staining tray. Using the wash bottle on the bench top, gently wash off the excess safranin from the slide., Place the slide on a staining tray and cover the entire film with methylene blue. Stain for one minute. step 3 Pick up the slide by one end and hold it at an angle over the staining tray. Using the wash bottle on the bench top, gently wash off the excess safranin from the slide., direct stain of Micrococcus luteus or Staphylococcus epidermidis step 1 Heat fix a smear of Micrococcus luteus or Staphylococcus epidermidis to a clean microscope slide., indirect staining done with acidic dye, basic dyes procedure direct stain of the normal flora and cells of your mouth, Aseptically remove a small amount of the culture from the agar surface and just touch it 2 - 3 times to the drop of water until it just turns cloudy. step 3 Burn the remaining bacteria off of the loop. If too much culture is added to the water, you will not see stained individual bacteria., acidic dye description The color portion of the dye resides in the negative ion., Heat fix a smear of Micrococcus luteus or Staphylococcus epidermidis to a clean microscope slide. step 2 Place the slide on a staining tray and cover the entire film with methylene blue. Stain for one minute., basic dyes procedure direct stain of Micrococcus luteus or Staphylococcus epidermidis, After the loop cools, spread the suspension over the entire slide to form a thin film. step 5 Allow this thin suspension to completely air dry., basic dyes description The color portion of the dye resides in the positive ion., Use a book of blotting paper to blot the slide dry. step 5 Focus using oil immersion microscopy. (1000X), Heat fix a smear of Escherichia coli or Enterobacter aerogenes to a clean microscope slide. step 2 Place the slide on a staining tray and cover the entire film with safranin. Stain for one minute., fixing the microbe to the slide step 1 Using the dropper bottle of distilled water, place 1/2 of a normal sized drop of water on a clean slide by touching the dropper to the slide, The color portion of the dye resides in the positive ion. mechanism of direct staining Because the cytoplasm of all bacterial cells has a slight negative charge, when using a basic dye, the positively charged color portion of the stain combines with the negatively charged bacterial cytoplasm (opposite charges attract) and the organism becomes directly stained., Pick up the slide by one end and hold it at an angle over the staining tray. Using the wash bottle on the bench top, gently wash off the excess safranin from the slide. step 4 Use a book of blotting paper to blot the slide dry., Using the dropper bottle of distilled water, place 1/2 of a normal sized drop of water on a clean slide by touching the dropper to the slide step 2 Aseptically remove a small amount of the culture from the agar surface and just touch it 2 - 3 times to the drop of water until it just turns cloudy., The color portion of the dye resides in the negative ion. mechanism of indirect staining Because the cytoplasm of all bacterial cells has a slight negative charge, when using an acidic dye, the negatively charged color portion of the stain is repelled by the negatively charged bacterial cytoplasm like charges repel and the dye forms a deposit around the organism. The background is stained, not the organism., direct staining preparing the slide for direct staining fixing the microbe to the slide, Place the slide on a staining tray and cover the entire film with safranin. Stain for one minute. step 3 Pick up the slide by one end and hold it at an angle over the staining tray. Using the wash bottle on the bench top, gently wash off the excess safranin from the slide.
