Confocal imaging of a gap junction protein associated with the keratocytes of the human cornea

Autor: Nigel H Brookes, CA Poole, GM Clover
Rok vydání: 1996
Předmět:
Zdroj: Australian and New Zealand Journal of Ophthalmology. 24:10-12
ISSN: 1440-1606
0814-9763
DOI: 10.1111/j.1442-9071.1996.tb00982.x
Popis: Keratocyte distribution and transparency make it difficult to investigate the precise microanatomy and intercellular connectivity of these cells throughout the cornea in siru. Earlier siudies were limited by the lack of resolution and the inability to achieve three-dimensional conceptualisation of keratocyte morphology.' More recently, human keratocytes have been visualised in situ using tandem scanning confocal microscopy in reflection mode, but resolution was again limited. Similarly, immunohistochemical staining of elements of the keratocyte cytoskeleton imaged by confocal laser scanning microscopy does not enable the keratocyte to be seen in its entirety.' Recently, our group reported the combined use of a new aldehyde fixable cell viability fluoroprobe, 5-chloromethylfluorescein diacetate (CMFDA), and confocal laser scanning microscopy to produce high-resolution images of the entire cytoplasm of the keratocyte ex uizo.''" Here we report on the co-localisation of an antibody to the gap junction protein connexin 50 on human corneal keratocytes stained with CMFDA. Dual channel confocal laser scanning microscopy, digital image reconstruction and merging techniques were used to produce highresolution images of connexin 50 distribution on the cell bodies and fine cell processes of mid-stroma1 keratocytes, and to highlight the functional potential of gap junction proteins in the maintenance of human corneal physiology.
Databáze: OpenAIRE
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