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Azim Akbarzadeh Dariush Norouzian Ali Farhangi Mohammad Reza Mehrabi Shirin Jamshidi Davood Zare Morvarid Shafiei 《Indian journal of clinical biochemistry : IJCB》2008,23(1):57-61
Flow cytometry has been employed as a method to study homogeneity of isolated islet subpopulations. After collagenase digestion
of rat pancreas and elutriation of tissue fragments, islets were isolated and dissociated, and cells were analyzed and sorted
according to their low forward angle light scattering properties by using automated flow cytometry. A standardized procedure
was developed for the preparation of rat islet cell grafts for purification of islet cells. In this process, after collagenase
digestion of pancreas, islets were isolated, dissociated, identification by dithizone method and then with enzymatic procedure
by DNase and trypsin, the islet cells changed into single cells and beta cells were identified by immunofluorescence method
and then assayed by flow cytometry. Methods have been developed for the preparation of suspension of viable rat pancreatic
islet cells and their analysis and sorting in the fluorescence activated cell sorter (FACC IV, Becton Dickinson, Sunnyvale,
Ca). Flow cytometry of these cells indicated that there were 91% of beta cells in cell suspension. Most of the exocrine particles
were lost during digestion. Purified endocrine islet cell grafts were prepared by pure beta-cells, without endocrine non-beta
cells. The purified aggregates were devoid of endocrine non-beta cells and damaged cells. 相似文献
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