Determine the average value for the epithelial cell centrosomes and multiply it by the normalization factor to determine the normalized value. Collect a Z stack and make a maximum intensity projection of the stack.Open the brightness and contrast window, select the color of the centrosome signal, and maximize both brightness and contrast to create a pseudobinary image of the centrosome.
Author information: (1)Shin Kong Wu Ho Su Memorial Hospital, Taipei, Taiwan. Unable to load your delegates due to an error If other labeling is present, go back Open the measurement window and select area measurement.Select the channel in which the centrosome signal was collected and adjust the slider down slightly so that everything but the centrosome signal is masked out.Using the overlay function, draw a circle 3 μm diameter around the centrosome.Select the area button to obtain the area of the centrosome signal inside the circle. For each tissue, calculate the average and standard deviation of the fibroblast centrosome size. Clipboard, Search History, and several other advanced features are temporarily unavailable. When added to unfixed cells, propidium iodide (PI) and ethidium bromide (EB) are taken up only by dying cells. It should be between 0.3 and 1.3 μmCollect a Z stack and make a maximum intensity projection of the stack.Open the brightness and contrast window, select the color of the centrosome signal, and maximize both brightness and contrast to create a pseudobinary image of the centrosome.Open the histogram window and select area measurement.Select the channel in which the centrosome signal was collected and adjust the low threshold slider down slightly so that everything but the centrosome signal is masked out.Using the overlay function, draw a circle ∼3 μm diameter around the first centrosome in the image.
ScienceDirect ® is a registered trademark of Elsevier B.V.URL: https://www.sciencedirect.com/science/article/pii/B9780123854933000024URL: https://www.sciencedirect.com/science/article/pii/B9780124160026000237URL: https://www.sciencedirect.com/science/article/pii/B9780124366435501218URL: https://www.sciencedirect.com/science/article/pii/S0076687900220442URL: https://www.sciencedirect.com/science/article/pii/S0076687906190072URL: https://www.sciencedirect.com/science/article/pii/S1874578405800170URL: https://www.sciencedirect.com/science/article/pii/B012227055X008361URL: https://www.sciencedirect.com/science/article/pii/B9780080885049002853URL: https://www.sciencedirect.com/science/article/pii/B9780120354177500088URL: https://www.sciencedirect.com/science/article/pii/B9780128018880000023A more objective method to determine if the cells expressing the gene of interest have indeed died (not just rounded up) is by differential uptake of fluorescent DNA-binding dyes. This method of SC isolation has been adapted for solid tissues such as the prostate ( 4 ) and, recently, unfractionated normal human renal tissue ( 15 ). The side population pattern is a result of both the nonlinear emission of Hoechst 33342 dye under UV excitation (Petersen et al., 2004) and the distinct ability of bone marrow cells to retain Hoechst dyes, and also likely the chromatin conformation within cell populations, which results in differential access of the dye to the DNA. A novel method that simultaneously monitors the low fluorescence intensity of Handbook of Immunohistochemistry and in situ Hybridization of Human Carcinomas, Volume 3Handbook of Immunohistochemistry and in Situ Hybridization of Human CarcinomasEncyclopedia of Food Sciences and Nutrition (Second Edition)From the onset of the use of flow cytometry for sex separation, using By staining intact cells with the DNA binding, vital, fluorescent dye, Pathological Assessment of Experimental Models of Stem Cell and Other Regenerative TherapiesStem Cell and Gene Therapy for Cardiovascular DiseaseScienceDirect ® is a registered trademark of Elsevier B.V. This site needs JavaScript to work properly. Tsai YL(1), Chang YJ, Chou CY, Cheong ML, Tsai MS.
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