Daniel:Notebook/HiResChrPaint/2013-11-29

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FISH Slide Prep[edit]

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I've got cells ready to split so I'm making some dishes for FISH.

FISH Slide Preparation[edit]

Original Protocol from Beliveau et al: File:Slide preparation for interphase FISH.pdf

  • Prepared 4 dishes PGP1F P9
1. Prepare a cell suspension of 500 000-2 000 000 cells/mL in growth media
2. Add 100 uL of cell suspension per dish roughly in the center
3. Allow cells to adhere for 3 hours at the 37 C
4. Rinse briefly in 1X PBS
5. Fix slides for 5-15 minutes in 4% (v/v) paraformaldehyde in 1X PBS
6. Rinse briefly in 1X PBS
7. Incubate for 5 minutes in 2X SSCT
8. Incubate for 5 minutes in 2X SSCT + 50% (v/v) formamide
9. Add fresh 2X SSCT + 50% (v/v) formamide for storage at 4C

Pictures[edit]

I took snapshots of all four dishes after fixing, just to be sure the cells were still there (since I've had trouble finding cells previously).


File:FISH 112913 dish11129 1752.jpg|File:FISH 112913 dish21129 1754.jpg|
Dish 1, 20X (left); Dish 2, 20X (right)
File:FISH 112913 dish31129 1755.jpg|File:FISH 112913 dish41129 1756.jpg|
Dish 3, 20X (left); Dish 4, 20X (right)