Table 1.
Synopsis of published protocols for quantitative oil red O staining of adipocytesa This table lists individual protocols for quantitative oil red O staining with as much detail as published, in chronological order of publication. A dash signifies that no information was given, which may indicate that a certain step was either not performed, or performed according to the base protocol, or performed differently but not reported. Protocols cited in the ‘based on’ column that are not explicitly listed are not quantitative protocols 24 or use a stain other than oil red O 25.
| First author, year | Based on | Wash | Fixate | Wash | Stain | Wash | Dry | Elute | Measure |
|---|---|---|---|---|---|---|---|---|---|
| Ramirez-Zacarias 1992 5 | 26 | — | 1 h RT 10% FA | water | 2 h RT 0.2% in 57% 2-prop | exhaustively | 32°C | 1 ml 2-prop | 510 nm |
| Bennett 2002 6 | 24 | — | — | — | — | — | — | 15 min 4% Igepalb | 520 nm |
| Janderova 2003 7 | 5 | — | 1 h RT 10% FA | 60% 2-prop | 10 min u/c in 60% 2-prop | water, repeatedly | — | 15 min 2-prop | 500 nm |
| Yang 2007 8 | — | 2x PBS | 1 h RT 10% FA | — | 1 h RT 0.6% in 60% 2-prop | 3x 60% 2-prop | — | 4% NP-40 in 2-prop | 490 nm |
| Mäuser 2007 9 | — | — | 15 min FA | — | 1 h 0.25% in 60% 2-prop | water, several times | — | 2-prop | 500 nm |
| Arsenescu 2008 10 | 5,25 | 1x PBS | 5 min RT 10% FA | — | 30 min 0.3% in 60% 2-prop | 2-prop | 510 nm | ||
| Rossmeislova 2013 11 | 7 | — | — | — | — | — | — | — | — |
| Wang 2015 12 | — | PBS | 30 min 4% FA | — | 20 min 0.3% | — | — | 2-prop | 518 nm |
| Cheung 2015 13 | — | — | — | — | — | — | — | 2-prop | 580 nm (sic) |
| Koc 2015 14 | 11 | — | — | — | — | — | — | — | — |
| Kim 2015 15 | —c | PBS | 1 h RT 10% FA | 2-prop | 2 h proprietary solutionc | — | — | 100% 2-prop | 520 nm |
| Mori 2016 18 | — | 2x PBS | 1 h RT 3.7% FA | — | 0.6% in 60% 2-prop | — | — | n/a | aread |
| Arbitrary protocol (Fig. 1) | 1x PBS | 15 min 4% FA | — | 60 min 0.3% in 40% 2-prop | water, 5 times | — | 10 min 100% 2-prop | 510 nm | |
| This protocol | 5 | 1x PBS | 15 min 4% FA | — | 30 min 0.132 ml/cm2 0.2% in 40% 2-prop | water, 5 times | — | 0.263 ml/cm2 100% 2-prop | 510 nm |
Note.
Abbreviations used in this table: 2-prop, 2-propanol; conc., concentration; FA, formaldehyde; n/a, not applicable; PBS, phosphate-buffered saline; RT, room temperature; u/c, unspecified concentration
4% Igepal CA-630 in 2-propanol
No academic article was cited, but a commercially available assay kit from Cayman Chemical; the actual protocol described in the article deviates substantially from the instructions provided by the manufacturer.
In this protocol, quantitative data was obtained by analyzing the percentage of stained area in microphotographs.