By L. Scott Cram (auth.), R. C. Sobti, Awtar Krishan (eds.)
Flow cytometry has quickly developed right into a procedure for speedy research of DNA content material, mobile marker expression and digital sorting of cells of curiosity for additional investigations. move cytometers are being largely used for tracking of mobile DNA content material, phenotype expression, drug delivery, calcium flux, proliferation and apoptosis. Phenotypic research of marker expression in leukemic cells has develop into a massive instrument for diagnostic and healing tracking of sufferers. fresh experiences have explored using stream cytometry for tracking hormone receptor expression in human stable tumors and for stories in human genomics. Contributions within the present quantity are in line with shows made on the First Indo-US workshop on stream Cytometry within which specialists from united states, united kingdom and India mentioned purposes of circulation cytometry in organic and scientific examine. This ebook can be of curiosity to submit graduates and researchers within the fields of pathology, cytology, cellphone biology and molecular biology.
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Additional info for Advanced Flow Cytometry: Applications in Biological Research
19. Wash the slide in a PBS solution well. 4. DNA staining 20. Treat the slides with RNase (l mg/mI, Sigma) for 30 minutes at room temperature to remove RNA from cells. 21. Transfer the slide to PBS solution containing propidium iodide (PI, 5 flg/ml, Calbiochem, La Jolla, CA). 22. Mount the slides in an anti-fade solution (Slow Fade, Molecular Probes) containing propidium iodide (PI, 5 flg/ml, Calbiochem). 23. Place coverslips on the slide. 24. Seal the slides with nail polish. , . , . ~"'.. 2.
Aspirate supernatant and flick tube to loosen pellet. 10. 0 ml of cold chromosome isolation buffer to each tube. Mix gently. 11. Vortex each tube vigorously for 30 sec. Place sample on ice for 10 min and observe chromosomes in microscope using propidium iodide to determine degree of dispersion. If needed, repeat vortexing for 10 sec and maintain sample on ice while rechecking dispersion. Repeat vortexing cycle as needed. 12. Chromosomes can be stained at this point or stored unstained at 4°C. 13.
5. Summary Flow cytogenetics is an exciting field that has made numerous contributions to our understanding of the human genome and karyotype instability. The results of these studies have been fruitful as they have been used to augment molecular, cytogenetic, and cell based investigations. The field continues to evolve into new and exciting areas as evidenced by the development of new applications for SNP detection and DNA fragment analysis. These assays will have an important role to play in understanding human heterogeneity and for the analysis of biological agents.
Advanced Flow Cytometry: Applications in Biological Research by L. Scott Cram (auth.), R. C. Sobti, Awtar Krishan (eds.)