Resolving morphology and antibody labeling over large distances in tissue sections. Microscopy research and technique

Marc D Jacobs, Paul J Donaldson, Mark B Cannell, Christian Soeller

Research output: Contribution to journalArticle (Academic Journal)

31 Citations (Scopus)

Abstract

Protein expression patterns are a primary determinant of tissue function and in this study we developed methods to study protein expression over macroscopic distances at subcellular levels of detail. Using the mammalian lens as a model tissue system, we show that by combining two-photon microscopy with novel image montage methods (fast beam blanking coupled with mathematical alignment tools) we have extended the limited field of view of laser scanning microscopes. To illustrate the utility of our approach, the distribution of connexin-46 was visualized across equatorial sections of the rat mammalian lens. By optimizing fixation protocols, good morphological preservation could be achieved over the thickness of the lens (approximately 4 mm) while preserving antigenicity of lens proteins. Using the same image data, changes in lens fiber cell morphology were mapped quantitatively by automatic image analysis routines. The methods presented should be generally applicable to any tissue system where changes in antibody labeling and tissue structure occur over large and small distances.
Original languageEnglish
Pages (from-to)83-91
Number of pages9
JournalMicroscopy Research and Technique
Volume62
Issue number1
Publication statusPublished - 1 Jan 2003

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