-
00:00
1.
Invasion Assay
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00:06
2.
Put transwell chambers in 24-well plate
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00:25
3.
Prepare Matrigel
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00:48
3.1
Add 100 ul Matrigel in the chamber
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01:09
4.
Incubate at 37 C overnight for gelling
-
01:15
5.
Add 750 ul culture medium with serum in the lower chamber
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01:26
6.
Place 200 ul cell (2.5x10^5/ml in serum-free medium) in upper chamber
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01:51
7.
Place transwell into lower chamber
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02:09
8.
Incubate at 37 C for 12-16 h
-
02:13
9.
Remove medium and wash twice by PBS
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02:52
10.
Fix cells by formaldehyde (3.7% in PBS)
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03:17
10.1
Fix cells at room temp for 2 min
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03:20
11.
Remove formaldehyde and wash twice by PBS
-
03:55
12.
Permeabilize cells by 100% methanol
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04:16
12.1
Permeabilization at room temp for 20 min
-
04:18
13.
Remove methanol and wash twice by PBS
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04:46
14.
Giemsa stain
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05:10
14.1
Giemsa stain at room temp for 15 min
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05:15
14.2
Remove Giemsa stain and wash twice by PBS
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05:43
15.
Scrape off non-invasive cells with cotton swabs
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06:04
16.
Count invasive cells under a light microscope