Bloodstain detection and discrimination from nonblood substances on various substrates are critical in forensic science as bloodstains are a critical source for confirmatory DNA tests. Conventional bloodstain detection methods often involve time-consuming sample preparation, a chance of harm to investigators, the possibility of destruction of blood samples, and acquisition of too little data at crime scenes either in the field or in the laboratory. An imaging method has the advantages of being nondestructive, noncontact, real-time, and covering a large field-of-view. The abundant spectral information provided by multispectral imaging makes it a potential presumptive bloodstain detection and discrimination method. This article proposes an interference filter (IF) based area scanning three-spectral-band crime scene imaging system used for forensic bloodstain detection and discrimination. The impact of large angle of views on the spectral shift of calibrated IFs is determined, for both detecting and discriminating bloodstains from visually similar substances on multiple substrates. Spectral features in the visible and near-infrared portion employed by the relative band depth method are used. This study shows that 1 ml bloodstain on black felt, gray felt, red felt, white cotton, white polyester, and raw wood can be detected. Bloodstains on the above substrates can be discriminated from cola, coffee, ketchup, orange juice, red wine, and green tea.