Activating long-lived room temperature phosphorescence (RTP) in the aqueous environment and thus realizing matrix-free, anti-oxygen, and time-resolved information encryption and cellular imaging remain a great challenge. Here, we fabricated three types of carbon dots (C-dots), i.e., fluorescent C-dots (F-C-dots) and two types of phosphorescent C-dots denoted as Pw-C-dots and Py-C-dots by a one-pot strategy. Their formation was attributed to the difference in the decarboxylation degree at high temperatures using trimesic acid (TMA) as a sole precursor. Unexpectedly, the yield reached as high as ∼92%, and the proportions were ∼27% for F-C-dots, ∼17% for Pw-C-dots, and ∼56% for Py-C-dots. These nanomaterials could help implement carbon peaking and carbon neutrality. Both green RTP of the two C-dots resulted from the small energy gap (ΔEST). These two RTP C-dots had a long lifetime of over 270 ms with a relatively high quantum yield (4.5 and 6.2%). They exhibited excellent photostability and anti-photobleaching performances. The dry and wet powders of the RTP C-dots were applied to high-level information encryption. The lifelike patterns were greatly different from those of the original ones and could last for several seconds to the naked eye, demonstrating that the RTP C-dots could be potentially employed as anti-oxygen and time-resolved contrast reagents. Most significantly, the cellular imaging experiments showed that the biofriendly PVP-coated Py-C-dots could localize at lysosomes and sustain hundreds of milliseconds. This approach not only pioneers a time-resolved lysosome localization model but also opens up a promising door for anti-oxygen and time-resolved RTP cytoimaging.