title: Single-step preparation and image-based counting of minute volumes of human blood.
abstract: Current flow-based blood counting devices require significant medical infrastructure and are not appropriate for field use. In this article we report on the development of a sample preparation, measurement, and analysis method that permits automated and accurate counting of red blood cells (RBCs), white blood cells (WBCs), and platelets, as well as allowing a 3-part differential of the WBCs to be performed on extremely small volumes of whole blood. This method is compatible with portable instrumentation that can be deployed in the field. The method consists of serially diluting blood samples first with sodium dodecyl sulfate dissolved in phosphate buffered saline, then in acridine orange dissolved in phosphate buffered saline, followed by fluorescence and dark field imaging with low magnification objectives. Image analysis is performed to extract cell counts and differentials. We performed a paired analysis of 20 volunteers with complete blood count values both within and beyond the normal reference range using a commercial automated hematology analyzer and the image-based method, with the new method achieving accuracies comparable to that of the commercial system. Because the sample preparation and imaging are simple and inexpensive to implement, this method has applications for pediatrics, clinician offices, and global health in regions that do not have access to central hematology laboratories.
authors: Center for Biophotonics Science and Technology, University of California, Davis, 2700 Stockton Blvd. Suite 1400, Sacramento, CA 95817, USA. swachsmann@ucdavis.edu.
authors: Zachary J
authors: ZJ
authors: Smith
authors: Tingjuan
authors: T
authors: Gao
authors: Kaiqin
authors: K
authors: Chu
authors: Stephen M
authors: SM
authors: Lane
authors: Dennis L
authors: DL
authors: Matthews
authors: Denis M
authors: DM
authors: Dwyre
authors: James
authors: J
authors: Hood
authors: Keith
authors: K
authors: Tatsukawa
authors: Laurence
authors: L
authors: Heifetz
authors: Sebastian
authors: S
authors: Wachsmann-Hogiu
mesh_terms: D000165:Acridine Orange / Q000737:chemistry; D001772:Blood Cell Count / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
chemical_list: D012967:Sodium Dodecyl Sulfate; D000165:Acridine Orange