1 00:00:04,000 --> 00:00:08,000 (English captions by Andrea Matsumoto, University of Michigan.) 2 00:00:15,000 --> 00:00:25,000 Prepare sputum smear by taking the sputum from the sputum container using the loop, 3 00:00:25,000 --> 00:00:30,000 which has been sterilized. 4 00:00:30,000 --> 00:00:39,000 Spread it on the microscope slide very wide, about two to three centimeter diameter and 5 00:00:39,000 --> 00:00:42,000 then flame the loop again. 6 00:00:42,000 --> 00:00:52,000 Flame the sputum smear by fixing it. 7 00:00:52,000 --> 00:01:11,000 These are the spirit lamp and then the stains required for the staining procedure. 8 00:01:11,000 --> 00:01:27,000 Cover the smear with Carbol Fuchsin. 9 00:01:27,000 --> 00:01:35,000 And heat it with the spirit lamp intermittently until it steams but should not boil or the 10 00:01:35,000 --> 00:02:03,000 steam should not get dried and should be done intermittently for five minutes. 11 00:02:03,000 --> 00:02:13,000 Tip the steam off and allow it to cool before washing the excess stain from the slide of 12 00:02:13,000 --> 00:02:21,000 the smear. 13 00:02:21,000 --> 00:02:46,000 Cover the smear with the decolorizer is twenty percent sulfuric acid for two minutes. 14 00:02:46,000 --> 00:02:55,000 After two minutes tip off the decolorizer and wash it under the tap water or distilled 15 00:02:55,000 --> 00:03:05,000 water. 16 00:03:05,000 --> 00:03:15,000 Cover the smear with the counterstain with methylene blue for two minutes. 17 00:03:15,000 --> 00:03:21,000 No heating. 18 00:03:21,000 --> 00:03:29,000 Wash it under the tap water or distilled water. 19 00:03:29,000 --> 00:03:39,000 And air dry it on the draining rack. 20 00:03:39,000 --> 00:03:50,000 Put oil immersion on the smeared slide and then examine it under the times hundred (100X) objective 21 00:03:50,000 --> 00:03:52,000 of the microscope. 22 00:03:52,000 --> 00:04:00,000 And you are expected to see reddish or pinkish bacilli, which can appear in a form of a cocci 23 00:04:00,000 --> 00:04:04,000 or it can also appear as a spiral or it can appear as a dot.