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

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.

journal_name

Microsc Res Tech

authors

Jacobs MD,Donaldson PJ,Cannell MB,Soeller C

doi

10.1002/jemt.10360

keywords:

subject

Has Abstract

pub_date

2003-09-01 00:00:00

pages

83-91

issue

1

eissn

1059-910X

issn

1097-0029

journal_volume

62

pub_type

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