[박상규] Oil Red O staining

[Anonymous]

                                                     2004.6.03                Arthur

Oil Red O staining

 

Prepare the following materials

: 0.5% Oil Red O stock Sol (in isopropanol), 10% formalin in PBS, Whatman #1 filter paper, glycerin jelly, 55 degree water bath

 

*Procedure

1.     Fix the cells or tissue in 10% formalin for 15 min or over night at room temperature

2.     Remove the formalin carefully and wash the sample with PBS, 2 times

3.   Prepare Working sol during the fixation (make fresh sol each time, working solution is very unstable) by adding 6ml stock sol to 4ml DW. Mix and filter through with whatman #1 filter paper (filtration step takes a while, DO Not hasten the filtration process by applying a vacuum because there is too much stuff to filter out)

4.     Apply 1.5ml working sol per one slide or 5ml per 100 mm dish

5.     Stand it for 1 hr at room temperature

6.     Carefully wash the slide or dish with DW

7.     Rinse several times with DW to remove excess staining or junk precipitate

8.     Maintain dish or slide in water if the intact cells are desired.

9.     Mount with 55 degree melted glycerin jelly (40ul/slide)

10.  watch

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