By Zbigniew Darzynkiewicz, John Paul Robinson, Harry A. Crissman, Leslie Wilson, Paul T. Matsudaira
Flow Cytometry, moment version offers an entire and accomplished quantity laboratory consultant and reference for using the most up-tp-date tools in movement cytometry pattern coaching and research. those crucial options are defined in a step by step structure, supplemented through explanatory sections and trouble-shooting assistance. The tools are obtainable to all researchers and scholars in biomedical technological know-how and biology who needs to use stream cytometry to split and examine cells. Key positive aspects * thoroughly revised and drastically accelerated because the e-book of the 1st variation in 1990 * equipment hide mobilephone loss of life and mobile cycle analyses useful, handbook-style presentation works in lab or school room * specified accomplished methodological insurance * colour plates illustrate concepts * In-depth remedy of strategies, together with an outline of every strategy: * Theoretical foundations * severe features * attainable pitfalls * Written via authors with vast event who: * constructed or changed the recommendations * Describe their event with varied tools and functions to various mobilephone structures * Are the who is Who in stream Cytometry
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Extra resources for Flow Cytometry Part A
Example text
Assays of Cell Viability bcl-2, c-myc, or tumor suppressor gene p53 and may be prognostic of treatment (Wyllie, 1992; Sachs and Lotem, 1993; Schwartzman and Cidlowski, 1993). Extensive research is underway in many laboratories to understand the mechanism of apoptosis. Knowledge of the molecular events of this process may be the basis for new antitumor strategies. , 1992). The most common feature of apoptosis is active participation of the cell in its self-annihilation. , 1990; Tomei and Cope, 1991; Compton, 1992).
Note presence of diploid range, cytokeratinnegative inflarnmatory/strornal cells. (d) Corresponding cytospin, showing intact tumor cells, bare nuclei, and host-derived inflammatory cells. 10 Daniel W. Visscher and John D. Crissman D. Morphologic Appearance of Suspensions We strongly recommend the use of cytospin preparations as a quality control measure in whole-cell dissociation procedures. In addition to reflecting neoplastic vs host cell representation, Papanicolaou-stained cytospin pellets also may be used to assess degree of cellular trauma/cytoplasmic stripping (see Fig.
488-nm laser line). Rh123 fluoresces green (530 20 nm), while PI, as described above, in red. * 23 2. Assays of Cell Viability .. n 0 20 40 60 80 0 20 40 60 80 0 20 40 60 80 I 100 Propidium Iodide Fig. 1 Stainability of live, apoptotic, and necrotic cells with Rh123 and PI. The mitochondria1 probe Rh123 (green fluorescence) is taken up by live cells from control HL-60 cultures (Control); live cells exclude PI. 15 yM camptothecin for 4 hr (CAM) neither changes Rh123 uptake nor affects exclusion of PI.