Microwave Digestion--Vacuum Filtration-Automated Scanning Electron Microscopy as a sensitive method for forensic diatom test.

Diagnosis of drowning is one of the most difficult issues in forensic practice. A number of methods have been developed over the years to determine whether a person was drowned. Microwave Digestion-Vacuum Filtration-Automated Scanning Electron Microscopy (MD-VF-Auto SEM) method we developed is a new qualitative and quantitative method of diatom test for diagnosis of drowning. The new method is based on microwave digestion technique, vacuum filtration, and automated SEM, which would achieve a maximal recovery of diatoms and identify diatoms easily by SEM with high resolution. This study was designed to evaluate the sensitivity of this method, the recovery of diatom, and loss ratio of centrifugation, which were compared using the MD-VF-Auto SEM method and the conventional acid digestion method. Two groups of samples were designed in the study. Groups A (n = 20) and B (n = 20) were performed by MD-VF-Auto SEM method and the conventional acid digestion method, respectively. In addition, another eight water samples were centrifuged, and the diatoms in the supernatant and precipitate were counted and measured, respectively, in order to find out how many diatoms were lost after centrifugation. The difference between the two groups was statistically highly significant, and about 34 % of diatoms were lost after centrifugation at 4,000 rpm for 15 min. The results showed that the MD-VF-Auto SEM method was more sensitive and specific.