Due to enormously attractive merits, such as outstanding chemical stability, excellent biocompatibility, unique photoluminescence characteristics, and salient sensing performances, carbon dots (CDs) present significant potentials in biochemical analysis. [9][10][11][12][13][14][15][16] In biochemical analysis, red fluorescence carbon dots (R-CDs, also red-emission carbon dots) hold better suitability owing to their desirable properties such as low scattering, [17][18][19][20] deep penetration, [21][22][23][24] and low phototoxicity to biological tissues. [25][26][27][28][29][30][31][32][33][34][35][36] Therefore, the development of R-CDs has been extensively explored in previous studies, such as those conducted by Yang and co-workers, [37] Xiong and co-workers, [34] and Yang et al. [35] R-CDs are generally obtained via two strategies: expansion of the π-conjugated (sp 2 hybridization) domain and incorporation of the n-orbital or impurity band between the π and π* orbitals. [38] The former usually uses conjugated molecules, such as benzenediol and its derivatives, [39] dihydroxynaphthalene, [40] and phloroglucinol, [41] as precursors; while the latter prefers to nonconjugated precursors with non-carbon heteroatoms, especially molecules including N, S, P moieties. [42] However, the both do not work with ideal effect. Conjugated molecules are rigid and limit their fusing, resulting in insufficient expansion of the π-conjugated domain; while heteroatoms hinder sp 2 graphite lattice formation. Consequently, new route to realize red emissive CDs still be explored. Among new efforts, an integrated solution based on the above two strategies, especially self-doped, is promising. Consequently, conjugated molecules with heteroatoms (N, S, P, and so on) are selected to synthesize R-CDs. Some molecules with small aromatic ring have been utilized to obtain R-CDs: such as phenylenediamine [43] and derivatives, [44] and other aromatic molecules with, NH 2 , NO 2 , COOH, NCO, or mixed functional groups. [26,45] A concise summary is available from the corresponding discussions in review article. [46] However, rigid polycyclic conjugated molecules (RPCMs) with multiple aromatic rings and substituted functional groups comprise large sp 2 domains, they are rarely utilized to prepare R-CDs, [47][48][49][50] due to the difficulties in fusion that originate from the steric hindrance. Although RPCM-based R-CDs have been demonstrated by Yuan et al., [51] Liu et al., [47] Li et al., [48] Jia et al., [49] Zheng et al., [50] and Tan et al., [45] suitability Red fluorescent carbon dots (R-CDs) are special desirable for biochemical analysis due to good biological compatibility and deep penetration; however, they remain as bottlenecks due to difficulties in expanding the sp 2 domain, especially those are fused from rigid polycyclic conjugated molecules (RPCMs) with heteroatom substituents due to huge steric hindrance and heteroatom blockage toward graphic lattice. Here, an RPCM with heteroatom substituents, 1,5-diamino-4,8-dihydroxyanthraq...