| CSX Transportation | Achan Subdivsion | 27.8926 | -81.97537 | 4 |
| CSX Transportation | Auburndale Subdivsion | 27.23065 | -81.00571 | 70 |
| CSX Transportation | Bainbridge Subdivision | 30.70491 | -86.01899 | 75 |
| CSX Transportation | Bone Valley Subdivison | 27.85947 | -81.96842 | 10 |
| CSX Transportation | Brewster Subdivision | 27.43538 | -81.88561 | 25 |
| CSX Transportation | Brooker Subdivision | 29.88363 | -82.33242 | 25 |
| CSX Transportation | Brookville Subdivision | 28.27799 | -82.55303 | 26 |
| CSX Transportation | Callahan Subdivision | 30.42865 | -81.93005 | 11 |
| CSX Transportation | Carters Subdivision | 28.16183 | -81.63018 | 20 |
| CSX Transportation | Clearwater Subdivision | 27.95096 | -82.71782 | 10 |
| CSX Transportation | Deerhaven Subdivision | 29.65049 | -82.32558 | 5 |
| CSX Transportation | Homestead Subdivision | 25.64175 | -80.364 | 20 |
| CSX Transportation | Jacksonville Terminal Subdivision | 30.34529 | -81.63545 | 25 |
| CSX Transportation | Kingsland Subdivision | 30.62867 | -81.6082 | 23 |
| CSX Transportation | Lakeland Subdivision | 28.01909 | -82.20872 | 16 |
| Norfolk Southern | Navair District | 30.41326 | -82.86526 | 20 |
| CSX Transportation | P&A Subdivision | 30.55796 | -85.80125 | 80 |
| CSX Transportation | Palmetto Subdivision | 27.58155 | -82.51183 | 26 |
| CSX Transportation | Plant City Subdivision | 28.01306 | -82.12034 | 5 |
| CSX Transportation | Sanford Subdivison | 29.38304 | -81.45967 | 72 |
| CSX Transportation | Tallahassee Subdivision | 30.39226 | -83.0272 | 80 |
| CSX Transportation | Tampa Terminal Subdivision | 27.95383 | -82.45146 | 10 |
| Norfolk Southern | Valdosta District | 30.50118 | -82.56028 | 55 |
| CSX Transportation | Valrico Subdivision | 27.8489 | -81.84216 | 10 |
| CSX Transportation | Vitis Subdivision | 28.24171 | -82.07787 | 20 |
| CSX Transportation | Wildwood Subdivision | 29.39218 | -82.09336 | 60 |
| CSX Transportation | Yeoman Subdivision | 28.08178 | -82.31732 | 14 |
| Florida East Coast Railway | Operations | 28.42213 | -78.74209 | 225 |
| Seminole Gulf Railway | Fort Myers and Sarasota Divisions | 26.46331 | -81.83117 | 15 |
| Florida Central Railroad | Operations | 28.754 | -81.808 | 27 |
| Bay Line Railroad | Operations | 30.8555 | -85.4387 | 53 |
| Florida Gulf and Atlantic Railroad | Operations | 30.58714 | -84.58325 | 175 |
| Amtrak | AutoTrain Operations | 28.80158 | -81.24291 | 5 |
| SunRail | Central Florida Rail Corridor | 28.52925 | -81.36925 | 35 |
| South Florida Regional Transportation Authority | South Florida Rail Corridor | 25.77427 | -80.19366 | 25 |
| South Florida Regional Transportation Authority | Tri-Rail | 26.27147 | -80.06737 | 37 |
| Florida Railroad Museum | Operations | 27.59 | -82.42484 | 15 |
| Gold Coast Railroad Museum | Operations | 25.61832 | -80.40298 | 2 |
| South Central Florida Express | Operations | 26.74478 | -80.92478 | 100 |
| Brightline | Operations | 27.99789 | -83.74395 | 300 |
| Apalachicola Northern Railroad | Operations | 27.99789 | -83.74395 | 300 |
