0:00:04.000,0:00:08.000 (English captions by Andrea Matsumoto, University of Michigan.) 0:00:15.000,0:00:25.000 Prepare sputum smear by taking the sputum[br]from the sputum container using the loop, 0:00:25.000,0:00:30.000 which has been sterilized. 0:00:30.000,0:00:39.000 Spread it on the microscope slide very wide,[br]about two to three centimeter diameter and 0:00:39.000,0:00:42.000 then flame the loop again. 0:00:42.000,0:00:52.000 Flame the sputum smear by fixing it. 0:00:52.000,0:01:11.000 These are the spirit lamp and then the stains[br]required for the staining procedure. 0:01:11.000,0:01:27.000 Cover the smear with Carbol Fuchsin. 0:01:27.000,0:01:35.000 And heat it with the spirit lamp intermittently[br]until it steams but should not boil or the 0:01:35.000,0:02:03.000 steam should not get dried and should be done[br]intermittently for five minutes. 0:02:03.000,0:02:13.000 Tip the steam off and allow it to cool before[br]washing the excess stain from the slide of 0:02:13.000,0:02:21.000 the smear. 0:02:21.000,0:02:46.000 Cover the smear with the decolorizer is twenty[br]percent sulfuric acid for two minutes. 0:02:46.000,0:02:55.000 After two minutes tip off the decolorizer[br]and wash it under the tap water or distilled 0:02:55.000,0:03:05.000 water. 0:03:05.000,0:03:15.000 Cover the smear with the counterstain with[br]methylene blue for two minutes. 0:03:15.000,0:03:21.000 No heating. 0:03:21.000,0:03:29.000 Wash it under the tap water or distilled water. 0:03:29.000,0:03:39.000 And air dry it on the draining rack. 0:03:39.000,0:03:50.000 Put oil immersion on the smeared slide and[br]then examine it under the times hundred (100X) objective 0:03:50.000,0:03:52.000 of the microscope. 0:03:52.000,0:04:00.000 And you are expected to see reddish or pinkish[br]bacilli, which can appear in a form of a cocci 0:04:00.000,0:04:04.000 or it can also appear as a spiral or it can[br]appear as a dot.