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In the first part of the study, reagents were standardized and validated in the BioMikro 200 blood counter (Bio-Medical Instruments). Flow cytometry and fluorescence microscopy were used to study the reagent performance. The reagents used were Wright staining solution (Rose Bengal, Hoechst 33342, Peridinin Chlorophyll, Rhodamine 123, AO/erythrocytes; Wright S) and Reagent Hematoxylin (Reag H ) (Sigma Chemical Co. Reagents). The fluorescent probes, when excited with violet light, showed increased fluorescence after staining with Wright S and Reag H. To validate the effectiveness of the Wright S solution and Reag H, unstained blood samples were stained with them and the number of cells was counted manually by the attending laboratory personnel. The result obtained was matched with the number obtained with the blood counter. The results demonstrated that the Wright S showed 2% decrease in cell count and Reag H showed 10% increase in cell count. The results were found to be quite similar to the cell count reported in the standard reference book.⁹

The System was used to initiate the infection cycle for the non human-primate use. The System was connected to an industrial centrifuge (Beckman Coulter GS-6R Centrifuge). The System was set to start the centrifuge when it was pre-programmed by the operator to start at 2, 4, 6, or 8 minutes from the end of the prescribed shake mode. Centrifugation at any time during the specified duration will cause the System to terminate the infection cycle and present the operator with a message “Oops, System error detected” indicating an error has been detected in the data acquisition system. If no error is detected by the operator then the system will restart the centrifuge if there was a power failure. The operator was instructed to place a stool sample in the stool collector vessel and initiate the shake/rotation sequence. He/she was not allowed to do anything else other than react to the error message. The operator is alerted to the error by the machine sound, a beep or even by the display of a message on the touch screen. The System operator was allowed to exit from the system and return to the procedure of attempting to obtain the appropriate buffy coat sample.

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