1 00:00:09,000 --> 00:00:13,000 (English captions by Andrea Matsumoto, University of Michigan.) 2 00:00:15,000 --> 00:00:20,000 These are the items we require in the stool concentration technique. 3 00:00:20,000 --> 00:00:27,000 We need the stool sample, the fine mesh of pore size three fifty to four fifteen micrometer, 4 00:00:27,000 --> 00:00:43,000 the fifteen mL centrifuge tube, a glass applicator for emulsifying the stool sample, the beaker 5 00:00:43,000 --> 00:00:58,000 for the filtrates, the ten percent Formol-saline for mixing the stool, then the diethyl ether. 6 00:00:58,000 --> 00:01:12,000 Add ten percent Formol-saline to the stool, about a gram of the stool sample. 7 00:01:12,000 --> 00:01:21,000 Emulsify the stool sample with the applicator. 8 00:01:21,000 --> 00:01:31,000 Emulsify it very well to get a smooth substance. 9 00:01:31,000 --> 00:01:53,000 Sift the stool sample through the fine mesh into the beaker. 10 00:01:53,000 --> 00:01:57,000 Discard the stool sample. 11 00:01:57,000 --> 00:02:03,000 The debris is discarded under the tap water because it has been disinfected already with 12 00:02:03,000 --> 00:02:07,000 the Formol. 13 00:02:07,000 --> 00:02:17,000 Transfer seven mils of stool filtrate into the fifteen mil centrifuge tube. 14 00:02:17,000 --> 00:02:30,000 Rinse whatever is left in the beaker into the centrifuge tube. 15 00:02:30,000 --> 00:02:40,000 Add three mils of the diethyl ether to the seven mil of the stool filtrate in the fifteen 16 00:02:40,000 --> 00:02:47,000 mil centrifuge tube making it to a total of ten mL. 17 00:02:47,000 --> 00:02:53,000 And we will see two layers: the ether layer and then the Formol-saline layer. 18 00:02:53,000 --> 00:03:06,000 Cover the centrifuge tube with a lid and mix the two layers very well. 19 00:03:06,000 --> 00:03:12,000 And thus the ether is to dissolve in the fat, which is present in the stool sample, to release 20 00:03:12,000 --> 00:03:16,000 the parasites. 21 00:03:16,000 --> 00:03:30,000 Put it in the centrifuge tube balancing it. 22 00:03:30,000 --> 00:03:40,000 Centrifuge at thousand five hundred (1500) RPM for two to five minutes. 23 00:03:40,000 --> 00:03:45,000 Okay take out the centrifuge tube and you will see four layers. 24 00:03:45,000 --> 00:03:51,000 The first layer is the ether layer, the debris layer, which is the insoluble pad, and then 25 00:03:51,000 --> 00:04:04,000 Formol-saline layer, and then the sediment which contains the parasite. 26 00:04:04,000 --> 00:04:17,000 Break through the debris layer and discard the first three layers under the tap water, 27 00:04:17,000 --> 00:04:22,000 which have already been treated with Formol. 28 00:04:22,000 --> 00:04:29,000 So the sediment will contain the parasites of the egg or the larvae and then cysts were 29 00:04:29,000 --> 00:04:36,000 suspended with part of the Formol-saline or saline. 30 00:04:36,000 --> 00:04:43,000 Use a disposable pipet to transmit it very well and transfer a drop onto the microscope 31 00:04:43,000 --> 00:04:45,000 slide. 32 00:04:45,000 --> 00:04:57,000 Cover the drop of smear with the cover slip and examine it under the microscope using 33 00:04:57,000 --> 00:05:01,000 the times ten objective (10X). 34 00:05:01,000 --> 00:05:05,000 Low light allows better to see the parasite of the egg, the cyst, and the larvae.