25 nmol of ZW800–1C was injected in 25 g CD-1 mice 4 h prior to imaging. (a) Representative color and NIR images of ZW800–1C showing major organs in the thoracic cavity (left), abdominal cavity (middle), and after resection (right). (b) Signal-to-background ratio (SBR) of resected major organs (Or) against muscle (Mu). Abbreviations used are He, heart; Lu, lung; Li, liver; Pa, pancreas; Sp, spleen; Ki, kidney; Du, duodenum; Ga, gallbladder; BD, bile duct; Bl, bladder; In, intestine. ***P = 0.0006 and ****P < 0.0001 by two-way ANOVA with Dunnett’s multiple comparison test. (c) Comparison of the pharmacokinetic parameters of the candidate NIR fluorophores. Abbreviations used are t1/2α, distribution blood half-life; t1/2β, elimination blood half-life; Vd, the volume of distribution; AUC, area under the curve; Cl, clearance. Data are shown as mean ± s.e.m.