The crude oils and their source rocks from Niger Delta basin, Nigeria were investigated by optical microscopy for the maceral compositions and by gas chromatography - mass spectrometry (GC-MS) for the composition, distribution and abundances of saturated and aromatic biomarkers. The data obtained showed that the rock samples and crude oils were formed from the mixed input of terrestrial and marine organic matter, and deposited under oxic to sub-oxic conditions in a lacustrine-fluvial/deltaic environment. The aliphatic maturity parameters calculated for the source rocks and oil samples showed that they have immature to early mature; and early oil window maturity respectively, whereas aromatic maturity ratios indicated that they were within immature to peak of oil generation and beyond; and peak to late oil generative window, respectively. The crude oils and source rocks were characterized by the dominance of C
2
-fluorenes over other alkylated homologues. The predominance of C
2
fluorenes over other alkylated homologues might have been influenced by source facies and depositional environments. The relative percentages of fluorenes (FLs %), dibenzofurans (DBFs %) and dibenzothiophenes (DBTs %) values for the rock samples range from 9.36 to 38.04 %, 35.82–71.60 % and 10.02–42.52 %, respectively, while the crude oils have ranges of values from 18.91 to 56.38 %, 24.90–72.34 % and 5.99–19.02 %, respectively. These values showed that the crude oils and source rocks originated from the mixture of organic matter (terrestrial and marine) with higher contribution from terrestrial organic matter and deposited in a marine/brackish-saline lacustrine/swampy environments. The abundance and distribution of fluorenes and its derivatives were found to be effective in determining the origin and depositional environments of crude oils and source rocks in the Niger Delta Basin, Nigeria.