دورية أكاديمية
Live-imaging of the Drosophila pupal eye.
| العنوان: | Live-imaging of the Drosophila pupal eye. |
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| المؤلفون: | Hellerman MB; Biology Department, Wesleyan University., Choe RH; Biology Department, Wesleyan University., Johnson RI; Biology Department, Wesleyan University; rijohnson@wesleyan.edu. |
| المصدر: | Journal of visualized experiments : JoVE [J Vis Exp] 2015 Jan 12 (95), pp. 52120. Date of Electronic Publication: 2015 Jan 12. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media |
| اللغة: | English |
| بيانات الدورية: | Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: | Drosophila/*cytology , Retina/*cytology, Animals ; Animals, Genetically Modified ; Drosophila Proteins/analysis ; Epithelium ; Female ; Green Fluorescent Proteins/analysis ; Male ; Pupa/cytology |
| مستخلص: | Inherent processes of Drosophila pupal development can shift and distort the eye epithelium in ways that make individual cell behavior difficult to track during live cell imaging. These processes include: retinal rotation, cell growth and organismal movement. Additionally, irregularities in the topology of the epithelium, including subtle bumps and folds often introduced as the pupa is prepared for imaging, make it challenging to acquire in-focus images of more than a few ommatidia in a single focal plane. The workflow outlined here remedies these issues, allowing easy analysis of cellular processes during Drosophila pupal eye development. Appropriately-staged pupae are arranged in an imaging rig that can be easily assembled in most laboratories. Ubiquitin-DE-Cadherin:GFP and GMR-GAL4-driven UAS-α-catenin:GFP are used to visualize cell boundaries in the eye epithelium (1-3). After deconvolution is applied to fluorescent images captured at multiple focal planes, maximum projection images are generated for each time point and enhanced using image editing software. Alignment algorithms are used to quickly stabilize superfluous motion, making individual cell behavior easier to track. |
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| المشرفين على المادة: | 0 (Drosophila Proteins) 147336-22-9 (Green Fluorescent Proteins) |
| تواريخ الأحداث: | Date Created: 20150205 Date Completed: 20160412 Latest Revision: 20181202 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC4354525 |
| DOI: | 10.3791/52120 |
| PMID: | 25651413 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-087X |
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| DOI: | 10.3791/52120 |