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Biomedical subjects

J G Atwood

Publications and source records attributed to J G Atwood.

4 recordsLinked to original sources

A high-performance system for automation of the polymerase chain reaction.

A high-performance PCR system has been developed which reduces the time required for PCR, increases the throughput, reduces reagent consumption and ensures reproducibility of amplification. Integration of sophisticated temperature control with optimally designed vessels has resulted in an amplification system which produces unique benefits. These include rapid amplification, the elimination of the need for oil, even for small volumes, and a microplate format which provides liquid handling automation benefits.

Automation↗

An evaluation of blood smears made by a new method using a spinner and diluted blood.

Blood smears were prepared with the use of a spinner, which rotated with a fixed velocity for a fixed time. All blood samples used for spun smears were diluted with a fixed ratio of buffered isotonic saline solution. Distribution of cells in these smears was found to be random. The average number of cells per unit area was substantially uniform from place to place on the same slide and on multiple slides made with the smae sample. The distribution of leukocytes by type was also iniform. For different blood samples, the average number of cells per unit area in the smears correlated well with the measured cell concentrations per unit volume in the samples for leukocytes, erythrocytes and platelets. Leukocyte differential counts on replicate spun smears using the same bloods also agreed to within the sampling error. They similarly agreed with differential counts on pulled smears made from undiluted samples of the same bloods. With few exceptions, erythrocytic morphology on the spun smears was comparable to that on the good areas of pulled smears made with undiluted samples of the same bloods. Nearly all the spun smears were suitable for both viual and fully automated hematologic examination for leukocytes, erythrocytes, and platelets. This was true over nearly the whole area of each spun slide. In these ways this spinner method makes smears whose consistently high quality is little affected by either the properties of the blood sample or the skill of maker.

Blood Cell Count↗

Making enzymatic methods optimum for measuring compounds with a kinetic analyzer.

Kinetic enzymatic methods for analysis of substrates can be made optimum for a sensitive photometric analyzer by adjusting the activity of the triggering (catalyzing) enzyme so that the reaction rate is maximum at the time of measurement. tat this optimum activity, the exponential time constant for exhaustion of substrate equals the time between triggering and rate measurement. The scale factor (defined as measured activity divided by sample concentration in the reaction mixture) is the same for all tests. Sensitivity to substrate concentration is predictable from instrumental absorbance uncertainty and molar absorptivity of the absorbing species. These predictions from Michaelis theory were verified experimentally for pyruvate and lactate triggered with lactate dehydrogenase, for glucose triggered with hexokinase, and for triglycerides triggered with glycerol kinase, the reaction rate being measured 30 s after triggering. Sensitivities of 1.5 times 10(-7) mol/liter were achieved. Serum diluted 1000-fold and analyzed for glucose gave a repeatability of 25 mg/liter with linearity to 4.0 g/liter. Samples diluted 300-fold and analyzed for triglycerides gave 30 mg/liter repeatability, with linearity to concentrations exceeding 3.0 g/liter.

Autoanalysis↗