Summary

कम्प्यूटर की सहायता विज़ुअलाइज़ेशन बड़े पैमाने पर और अग्नाशय आइलेट मास, Size वितरण और वास्तुकला की मात्रा

Published: March 04, 2011
doi:

Summary

(1) पूरे खंड के आभासी स्लाइस पर कब्जा, (2) बड़े पैमाने पर डेटा के मास विश्लेषण, (3) 2 डी आभासी स्लाइस के पुनर्निर्माण: उपन्यास बड़े पैमाने पर और immunohistochemically दाग अग्नाशय के नमूनों की खरीद विश्लेषण के कंप्यूटर की सहायता तरीकों में वर्णित हैं 3D आइलेट मानचित्रण (4), और (5) गणितीय विश्लेषण.

Abstract

The pancreatic islet is a unique micro-organ composed of several hormone secreting endocrine cells such as beta-cells (insulin), alpha-cells (glucagon), and delta-cells (somatostatin) that are embedded in the exocrine tissues and comprise 1-2% of the entire pancreas. There is a close correlation between body and pancreas weight. Total beta-cell mass also increases proportionately to compensate for the demand for insulin in the body. What escapes this proportionate expansion is the size distribution of islets. Large animals such as humans share similar islet size distributions with mice, suggesting that this micro-organ has a certain size limit to be functional. The inability of large animal pancreata to generate proportionately larger islets is compensated for by an increase in the number of islets and by an increase in the proportion of larger islets in their overall islet size distribution. Furthermore, islets exhibit a striking plasticity in cellular composition and architecture among different species and also within the same species under various pathophysiological conditions. In the present study, we describe novel approaches for the analysis of biological image data in order to facilitate the automation of analytic processes, which allow for the analysis of large and heterogeneous data collections in the study of such dynamic biological processes and complex structures. Such studies have been hampered due to technical difficulties of unbiased sampling and generating large-scale data sets to precisely capture the complexity of biological processes of islet biology. Here we show methods to collect unbiased “representative” data within the limited availability of samples (or to minimize the sample collection) and the standard experimental settings, and to precisely analyze the complex three-dimensional structure of the islet. Computer-assisted automation allows for the collection and analysis of large-scale data sets and also assures unbiased interpretation of the data. Furthermore, the precise quantification of islet size distribution and spatial coordinates (i.e. X, Y, Z-positions) not only leads to an accurate visualization of pancreatic islet structure and composition, but also allows us to identify patterns during development and adaptation to altering conditions through mathematical modeling. The methods developed in this study are applicable to studies of many other systems and organisms as well.

Protocol

1. Immunohistochemically सना हुआ छवियाँ के आभासी स्लाइस बनाना ओपन StereoInvestigator. साफ नमूना खुर्दबीन धारक में पकड़े स्लाइड प्लेस और स्टीरियो अन्वेषक में यह "अधिग्रहण" पर क्लिक करके और फिर "लाइव छवि" (अधिग्रहण → लाइ?…

Discussion

कंप्यूटर की मदद से बड़े पैमाने पर दृश्य और मात्रा का ठहराव यहाँ प्रस्तुत अग्नाशय आइलेट अध्ययन में चार महत्वपूर्ण बिंदुओं को वहन: (1) अग्नाशय के नमूनों की बड़े पैमाने पर विश्लेषण समग्र आइलेट आकार के वित?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

अध्ययन के द्वारा समर्थित है अमेरिका के सार्वजनिक स्वास्थ्य सेवा डी.के.-081,527 अनुदान, डी.के. – ०,७२,४७३ और शिकागो मधुमेह अनुसंधान और प्रशिक्षण केन्द्र (पशु मॉडल कोर) के विश्वविद्यालय, और Kovler परिवार फाउंडेशन से एक उपहार के लिए डी.के.-+२०,५९५.

Materials

Material Name Type Company Catalogue Number Comment
Fluorescent microscope Microscope Olympus IX-81  
Stereo Investigator Program MicroBrightField    
MIP-GFP mice Mice Jackson Laboratory    
Mathematica Program Wolfram    
Image J Program NIH    
Slidebook   Program Olympus  

References

  1. Steiner, D. J., Kim, A., Miller, K., Hara, M. Pancreatic islet plasticity – Interspecies comparison of islet architecture and composition. ISLETS. 2, 135-145 (2010).
  2. Kim, A., Miller, K., Jo, J., Wojcik, P. l., Kilimnik, G., Hara, M. Islet architecture – a comparative study. ISLETS. 1, 129-136 (2009).
  3. Kilimnik, G., Kim, A., Jo, J., Miller, K., Hara, M. Quantification of pancreatic islet distribution in situ in mice. Am J Physiol Endocrinol Metab. 297, E1331-E1338 (2009).
  4. Hara, M., Dizon, R. F., Glick, B. S., Lee, C. S., Kaestner, K. H., Piston, D. W., Bindokas, V. P. Imaging pancreatic beta-cells in the intact pancreas. Am J Physiol Endocrinol Metab. 290, E1041-E1047 (2006).
  5. Hara, M., Wang, X., Kawamura, T., Bindokas, V. P., Dizon, R. F., Alcoser, S. Y., Magnuson, M. A., Bell, G. I. Transgenic mice with green fluorescent protein-labeled pancreatic beta -cells. Am J Physiol Endocrinol Metab. 284, E177-E183 (2003).
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Cite This Article
Kim, A., Kilimnik, G., Guo, C., Sung, J., Jo, J., Periwal, V., Witkowski, P., Dilorio, P., Hara, M. Computer-assisted Large-scale Visualization and Quantification of Pancreatic Islet Mass, Size Distribution and Architecture. J. Vis. Exp. (49), e2471, doi:10.3791/2471 (2011).

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