[Study on the standardization of chemical autoanalysis. Blood sugar. 3].
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A semiautomatic method for sperm analysis is described. The method combines a previously developed multiple exposure photography system with a digitizer connected to a computer. An undiluted specimen is inserted into a 10-mic chamber and the sperms are microscopically photographed for 1 second while illuminated by six short light pulses. On the developed film nonmotile sperms appear very bright, while motile sperms are seen as six-ringed chains. In addition, high magnification photographs of immobilized preheated sperms are taken for morphology analysis. The photographed images are projected onto sheets of paper where the traces of the nonmotile and motile sperms as well as the normal and abnormal forms are marked by a pen. For motility determination, sheets containing the sperm tracks are placed on the digitizer tablet. By means of a sonic pen the coordinates of the ends of the tracks are automatically recorded and the number of motile and nonmotile sperms are counted. Then the percent of abnormal forms are entered at the keyboard. Within a few seconds the computer supplies a printout of the results, including sperm concentration, percent of motile sperms, average velocity and frequency distribution of velocities of moving sperms, percent of abnormal forms, and total number of analyzed sperms.
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Reproducibility of results and possible clinical use of a newly developed noncontact autofocus specular microscope were studied. Morphological parameters of corneal endothelium were analyzed on images taken with the noncontact autofocus specular microscope and the ordinary contact specular microscope simultaneously in 10 eyes of 10 normal subjects. Intraphotographic and interphotographic study by autoanalysis of 5 images each of 10 eyes taken by the noncontact instrument showed small coefficient variation of mean cell area and cell density (less than 5%, individually). These morphological parameters showed about 7% difference between autoanalysis and manual analysis of the images taken by the noncontact instrument. In comparison with the images taken by the contact instrument, the images taken by the noncontact instrument showed about 7% difference in mean cell area and cell density (p < 0.001, individually). In respect of reproducibility, this newly developed noncontact autofocus specular microscope is clinically useful. The differences of morphological parameters between the new and ordinary instruments might be caused the differences in corneal shapes.
AutoAnalysis is a new method for detecting and quantitating the trityl cation released each cycle on automated DNA synthesizers. The trityl (dimethoxytrityl) cation is removed from the growing oligonucleotide after each base addition and is a useful measure of synthesis efficiency. The traditional absorbance method of collecting each trityl effluent with a fraction collector, followed by dilution with an acid solution and careful quantitation by UV/VIS spectroscopy is costly, tedious and prone to error. The absorbance method for trityl cation analysis must usually wait until the synthesis is complete. Interruption of a failed operation, for a variety of reasons, such as an empty reagent reservoir, is thus not possible. Taking advantage of the conductive properties of the trityl cation, immediate and real-time quantitation is now possible by integrating the total conductance of the flowing stream during the detritylation step after each nucleoside addition in DNA synthesis. A conductivity cell is mounted downstream, past the synthesis column. The conductivity signal is processed and displayed as the current average stepwise yield and overall yield. If the yield drops below a pre-set threshold value because of a failure situation, the synthesizer will interrupt, preserving reagents. AutoAnalysis allows trityl monitoring with complete automation on the Applied Biosystems Models 392 and 394 DNA/RNA Synthesizers.
The authors studied comparatively the sensitivity of 3 bacterial strains (P. Aeruginosa and two E. coli) with regard to antibiotics using an automatic method (ABAC) compared with a gel diffusion technic with impregnated disks. Each strain was tested by both methods 50 to 55 times. The diffusion method was easily reproducible with an average standard deviation of 2.23 mm. Its results were in agreement compared with the average diameter of measurements in 93.3% of cases. The automatic method gave results which agreed together in 95.7% of cases. It is thus reproducible. If one compares it with the results of the diffusion method, considered here as the reference method, we obtained the same response in 86.3% of cases. In 13.7% there was disagreement in interpretation (in most cases, the strain found intermediate in gel diffusion was considered sensitive on autoanalysis).