[Script Info]
Title: 
[Events]
Format: Layer, Start, End, Style, Name, MarginL, MarginR, MarginV, Effect, Text
Dialogue: 0,0:00:03.00,0:00:11.00,Default,,0000,0000,0000,,(English captions by Andrea Matsumoto, University of Michigan.) [music]
Dialogue: 0,0:00:11.00,0:00:21.00,Default,,0000,0000,0000,,Step 1. Prepare the smear
Dialogue: 0,0:00:21.00,0:00:28.00,Default,,0000,0000,0000,,Prepare the smear by flaming the loop taking\Nyour inoculum from a fresh culture.
Dialogue: 0,0:00:28.00,0:00:35.00,Default,,0000,0000,0000,,After you have put on a drop of sealant on\Na slide.
Dialogue: 0,0:00:35.00,0:00:40.00,Default,,0000,0000,0000,,Take a culture, dip part of the inoculum \Nin the culture.
Dialogue: 0,0:00:40.00,0:00:47.00,Default,,0000,0000,0000,,Take the culture and moistify it on the slide\Nto join with the rest of the sealant to make
Dialogue: 0,0:00:47.00,0:00:54.00,Default,,0000,0000,0000,,a smear of about two to three centimeters.
Dialogue: 0,0:00:54.00,0:00:59.00,Default,,0000,0000,0000,,Step 2. Gently heat fix the slide (after it\Nhas air-dried.)
Dialogue: 0,0:00:59.00,0:01:05.00,Default,,0000,0000,0000,,So, after air-drying, heat fix the smear by\Npassing it through the flame about three or
Dialogue: 0,0:01:05.00,0:01:07.00,Default,,0000,0000,0000,,four times.
Dialogue: 0,0:01:07.00,0:01:12.00,Default,,0000,0000,0000,,This makes the organism to die or kill the\Norganism.
Dialogue: 0,0:01:12.00,0:01:18.00,Default,,0000,0000,0000,,Allows you to stick onto the slide and then\Nprevents autolysis and makes the organism
Dialogue: 0,0:01:18.00,0:01:21.00,Default,,0000,0000,0000,,permeable to the stain.
Dialogue: 0,0:01:21.00,0:01:25.00,Default,,0000,0000,0000,,Step 3. Flood the smear with crystal violet
Dialogue: 0,0:01:25.00,0:01:36.00,Default,,0000,0000,0000,,Flood the smear with crystal violet for one\Nminute.
Dialogue: 0,0:01:36.00,0:01:42.00,Default,,0000,0000,0000,,Allow it to stain for 60 seconds.
Dialogue: 0,0:01:42.00,0:01:58.00,Default,,0000,0000,0000,,Tip off the excess crystal violet stain and\Nwash under gentle tap water.
Dialogue: 0,0:01:58.00,0:02:05.00,Default,,0000,0000,0000,,Step 4. Flood the slide with Lugol's iodine\Nsolution
Dialogue: 0,0:02:05.00,0:02:14.00,Default,,0000,0000,0000,,Cover the smear with Lugol's iodine for\None minute.
Dialogue: 0,0:02:14.00,0:02:18.00,Default,,0000,0000,0000,,Allow it to stain for 60 seconds.
Dialogue: 0,0:02:18.00,0:02:22.00,Default,,0000,0000,0000,,Wash off the excess Lugol's iodine stain\Nwith gentle tap water.
Dialogue: 0,0:02:22.00,0:02:31.00,Default,,0000,0000,0000,,Step 5. Decolorize briefly with acetone.
Dialogue: 0,0:02:31.00,0:02:40.00,Default,,0000,0000,0000,,Decolorize the smear with acetone for ten\Nseconds.
Dialogue: 0,0:02:40.00,0:02:52.00,Default,,0000,0000,0000,,Tip off the excess alcohol and wash under\Ntap water, gentle tap water.
Dialogue: 0,0:02:52.00,0:02:58.00,Default,,0000,0000,0000,,Step 6. Counterstain with neutral red (or\Nsafranin).
Dialogue: 0,0:02:58.00,0:03:03.00,Default,,0000,0000,0000,,Use neutral red to counter-stain for one minute.
Dialogue: 0,0:03:03.00,0:03:07.00,Default,,0000,0000,0000,,Cover the smear with the neutral red.
Dialogue: 0,0:03:07.00,0:03:14.00,Default,,0000,0000,0000,,Allow it to stain for 60 seconds.
Dialogue: 0,0:03:14.00,0:03:26.00,Default,,0000,0000,0000,,Wash off the excess neutral red under tap\Nwater.
Dialogue: 0,0:03:26.00,0:03:31.00,Default,,0000,0000,0000,,And blot it dry with a filter paper or a blotting\Npaper.
Dialogue: 0,0:03:31.00,0:03:36.00,Default,,0000,0000,0000,,Don't wipe.
Dialogue: 0,0:03:36.00,0:03:48.00,Default,,0000,0000,0000,,The slide is now ready to be examined microscopically\Nat 100X (oil immersion).
Dialogue: 0,0:03:48.00,0:04:01.00,Default,,0000,0000,0000,,Put oil immersion on the slide where the smear\Nis.
Dialogue: 0,0:04:01.00,0:04:04.00,Default,,0000,0000,0000,,And put the slide under the microscope.
Dialogue: 0,0:04:04.00,0:04:08.00,Default,,0000,0000,0000,,We are using the five hundred objective (500X\Nzoom). Examine the slide.