-
00:06
1.
Invasion Assay
-
00:19
2.
Put transwell chambers in 24-well plate
-
00:22
3.
Prepare Matrigel
-
00:21
3.1
Add 100 ul Matrigel in the chamber
-
00:06
4.
Incubate at 37 C overnight for gelling
-
00:11
5.
Add 750 ul culture medium with serum in the lower chamber
-
00:24
6.
Place 200 ul cell (2.5x10^5/ml in serum-free medium) in upper chamber
-
00:18
7.
Place transwell into lower chamber
-
00:03
8.
Incubate at 37 C for 12-16 h
-
00:39
9.
Remove medium and wash twice by PBS
-
00:25
10.
Fix cells by formaldehyde (3.7% in PBS)
-
00:03
10.1
Fix cells at room temp for 2 min
-
00:34
11.
Remove formaldehyde and wash twice by PBS
-
00:20
12.
Permeabilize cells by 100% methanol
-
00:02
12.1
Permeabilization at room temp for 20 min
-
00:28
13.
Remove methanol and wash twice by PBS
-
00:23
14.
Giemsa stain
-
00:04
14.1
Giemsa stain at room temp for 15 min
-
00:28
14.2
Remove Giemsa stain and wash twice by PBS
-
00:20
15.
Scrape off non-invasive cells with cotton swabs
-
00:12
16.
Count invasive cells under a light microscope