Staining techniques controls

Kapp et al. (1979) reported the presence of Y-chromosomal non-disjunction in 1,2-dibromo-3-chloropropane-cxposed workers using a quinacrine-staining technique. [There was no indication of the level of exposure to 1,2-dibromo-3-chloropropane or whether other exposures were present.] The frequency of sperm with two spots (indicating two Y chromosomes) was 1.2% (range, 0.8-1.8%) in 15 controls and 3.8% (range, 2.0-5.3%) in 18 samples from exposed men. 

I will first describe leukocytes in a general way for those not familiar with this family of particles. We will then proceed to a description of general staining techniques for surface markers, with attention to the need for controls. At this point I will digress slightly and discuss the way cytometry results can (and cannot) be quantified and the way sensitivity can affect these results. Finally, I will discuss gating strategies, in both philosophy and practice. 

These are indicative of proper staining techniques and provide a measure of whether the target retrieval procedure has been carried out correctly. They should assess correct temperature and incubation period of water baths or other retrieval methods. Likewise, positive tissue controls verify that all reagents were applied, that they performed correctly, and the proper incubation time and temperature were used. 

Hb is another analyte, which is easily amenable to CE analysis. This is because of its high clinical concentrations as well as its homogeneity. UV detection (absorption of 415 nm visible light) of Hb is more accurate than the dye-staining technique for protein quantitation. Hb variants such as HbS, HbC, and HbA (normal control) were subjected to CE analysis. With an optimized running buffer, the three species of Hbs were separated as three distinct peaks in the electropherogram (Fig. 3a) these results are comparable to conventional gel electrophoresis (Fig. 3b). >... 

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