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Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy
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| Zeitschriftentitel: | Microcirculation |
|---|---|
| Personen und Körperschaften: | , |
| In: | Microcirculation, 2, 1995, 3, S. 267-276 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
|
| Schlagwörter: |
| author_facet |
He, P. Adamson, R. H. He, P. Adamson, R. H. |
|---|---|
| author |
He, P. Adamson, R. H. |
| spellingShingle |
He, P. Adamson, R. H. Microcirculation Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy Physiology (medical) Cardiology and Cardiovascular Medicine Molecular Biology Physiology |
| author_sort |
he, p. |
| spelling |
He, P. Adamson, R. H. 1073-9688 1549-8719 Wiley Physiology (medical) Cardiology and Cardiovascular Medicine Molecular Biology Physiology http://dx.doi.org/10.3109/10739689509146772 <jats:title>ABSTRACT</jats:title><jats:p><jats:bold>Objective:</jats:bold> The cellular basis for spatial heterogeneity along postcapillary venules in their response to inflammatory mediators is not understood. To study permeability regulatory processes on an individual cell basis in intact microvessels, we developed methods to delineate individual endothelial cells within living, single perfused microvessels.</jats:p><jats:p><jats:bold>Methods:</jats:bold> Individual postcapillary microvessels in the mesenteries of frogs and hamsters were perfused with chloride‐free Ringer's solutions containing AgNO<jats:sub>3</jats:sub> (0.1 g per 100 ml) for 5–10 s. Vessels were immediately flushed with Ringer's solution containing serum albumin and some vessels subsequently perfused with the fluorescent nucleic acid stain YO‐PR0‐1 (1 μM) for 10–15 min. Vessels were imaged in situ using laser scanning confocal microscopy. A reflectance image from silver precipitate in the endothelial clefts and a fluorescence image of the nuclei were simultaneously recorded. Stacks of confocal images were merged and used to reconstruct the three‐dimensional orientation of endothelial cells. Hydraulic conductivity of some frog mesenteric venular vessels, measured before and after perfusion with AgNO<jats:sub>3</jats:sub>, was used to assess the integrity of the vessels.</jats:p><jats:p><jats:bold>Results:</jats:bold> The endothelial cell clefts were delineated with a very fine, readily imaged precipitate of silver. Hydraulic conductivity measured after AgNO<jats:sub>3</jats:sub> perfusion in frog mesenteric venular microvessels was not significantly different from control. Vessels showed a slight reduction in reactivity to inflammatory stimuli. The three‐dimensional pattern of endothelial cells in the living vessels was imaged.</jats:p><jats:p><jats:bold>Conclusions:</jats:bold> Combining confocal microscopy with silver staining in single perfused microvessels was shown to be an effective means to delineate the three‐dimensional pattern of endothelial cells forming microvessel walls. It enables further study of the vascular structure and function relationships at the individual cell level in intact microvessels.</jats:p> Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy Microcirculation |
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10.3109/10739689509146772 |
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Biologie Medizin |
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Wiley, 1995 |
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Wiley, 1995 |
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1073-9688 1549-8719 |
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he1995visualizationofendothelialcleftsandnucleiinlivingmicrovesselswithcombinedreflectanceandfluorescenceconfocalmicroscopy |
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1995 |
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Microcirculation |
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49 |
| title |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_unstemmed |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_full |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_fullStr |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_full_unstemmed |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_short |
Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_sort |
visualization of endothelial clefts and nuclei in living microvessels with combined reflectance and fluorescence confocal microscopy |
| topic |
Physiology (medical) Cardiology and Cardiovascular Medicine Molecular Biology Physiology |
| url |
http://dx.doi.org/10.3109/10739689509146772 |
| publishDate |
1995 |
| physical |
267-276 |
| description |
<jats:title>ABSTRACT</jats:title><jats:p><jats:bold>Objective:</jats:bold> The cellular basis for spatial heterogeneity along postcapillary venules in their response to inflammatory mediators is not understood. To study permeability regulatory processes on an individual cell basis in intact microvessels, we developed methods to delineate individual endothelial cells within living, single perfused microvessels.</jats:p><jats:p><jats:bold>Methods:</jats:bold> Individual postcapillary microvessels in the mesenteries of frogs and hamsters were perfused with chloride‐free Ringer's solutions containing AgNO<jats:sub>3</jats:sub> (0.1 g per 100 ml) for 5–10 s. Vessels were immediately flushed with Ringer's solution containing serum albumin and some vessels subsequently perfused with the fluorescent nucleic acid stain YO‐PR0‐1 (1 μM) for 10–15 min. Vessels were imaged in situ using laser scanning confocal microscopy. A reflectance image from silver precipitate in the endothelial clefts and a fluorescence image of the nuclei were simultaneously recorded. Stacks of confocal images were merged and used to reconstruct the three‐dimensional orientation of endothelial cells. Hydraulic conductivity of some frog mesenteric venular vessels, measured before and after perfusion with AgNO<jats:sub>3</jats:sub>, was used to assess the integrity of the vessels.</jats:p><jats:p><jats:bold>Results:</jats:bold> The endothelial cell clefts were delineated with a very fine, readily imaged precipitate of silver. Hydraulic conductivity measured after AgNO<jats:sub>3</jats:sub> perfusion in frog mesenteric venular microvessels was not significantly different from control. Vessels showed a slight reduction in reactivity to inflammatory stimuli. The three‐dimensional pattern of endothelial cells in the living vessels was imaged.</jats:p><jats:p><jats:bold>Conclusions:</jats:bold> Combining confocal microscopy with silver staining in single perfused microvessels was shown to be an effective means to delineate the three‐dimensional pattern of endothelial cells forming microvessel walls. It enables further study of the vascular structure and function relationships at the individual cell level in intact microvessels.</jats:p> |
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| author | He, P., Adamson, R. H. |
| author_facet | He, P., Adamson, R. H., He, P., Adamson, R. H. |
| author_sort | he, p. |
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| description | <jats:title>ABSTRACT</jats:title><jats:p><jats:bold>Objective:</jats:bold> The cellular basis for spatial heterogeneity along postcapillary venules in their response to inflammatory mediators is not understood. To study permeability regulatory processes on an individual cell basis in intact microvessels, we developed methods to delineate individual endothelial cells within living, single perfused microvessels.</jats:p><jats:p><jats:bold>Methods:</jats:bold> Individual postcapillary microvessels in the mesenteries of frogs and hamsters were perfused with chloride‐free Ringer's solutions containing AgNO<jats:sub>3</jats:sub> (0.1 g per 100 ml) for 5–10 s. Vessels were immediately flushed with Ringer's solution containing serum albumin and some vessels subsequently perfused with the fluorescent nucleic acid stain YO‐PR0‐1 (1 μM) for 10–15 min. Vessels were imaged in situ using laser scanning confocal microscopy. A reflectance image from silver precipitate in the endothelial clefts and a fluorescence image of the nuclei were simultaneously recorded. Stacks of confocal images were merged and used to reconstruct the three‐dimensional orientation of endothelial cells. Hydraulic conductivity of some frog mesenteric venular vessels, measured before and after perfusion with AgNO<jats:sub>3</jats:sub>, was used to assess the integrity of the vessels.</jats:p><jats:p><jats:bold>Results:</jats:bold> The endothelial cell clefts were delineated with a very fine, readily imaged precipitate of silver. Hydraulic conductivity measured after AgNO<jats:sub>3</jats:sub> perfusion in frog mesenteric venular microvessels was not significantly different from control. Vessels showed a slight reduction in reactivity to inflammatory stimuli. The three‐dimensional pattern of endothelial cells in the living vessels was imaged.</jats:p><jats:p><jats:bold>Conclusions:</jats:bold> Combining confocal microscopy with silver staining in single perfused microvessels was shown to be an effective means to delineate the three‐dimensional pattern of endothelial cells forming microvessel walls. It enables further study of the vascular structure and function relationships at the individual cell level in intact microvessels.</jats:p> |
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| imprint | Wiley, 1995 |
| imprint_str_mv | Wiley, 1995 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-D161, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-14, DE-Ch1, DE-L229 |
| issn | 1073-9688, 1549-8719 |
| issn_str_mv | 1073-9688, 1549-8719 |
| language | English |
| last_indexed | 2024-03-01T15:06:54.417Z |
| match_str | he1995visualizationofendothelialcleftsandnucleiinlivingmicrovesselswithcombinedreflectanceandfluorescenceconfocalmicroscopy |
| mega_collection | Wiley (CrossRef) |
| physical | 267-276 |
| publishDate | 1995 |
| publishDateSort | 1995 |
| publisher | Wiley |
| record_format | ai |
| recordtype | ai |
| series | Microcirculation |
| source_id | 49 |
| spelling | He, P. Adamson, R. H. 1073-9688 1549-8719 Wiley Physiology (medical) Cardiology and Cardiovascular Medicine Molecular Biology Physiology http://dx.doi.org/10.3109/10739689509146772 <jats:title>ABSTRACT</jats:title><jats:p><jats:bold>Objective:</jats:bold> The cellular basis for spatial heterogeneity along postcapillary venules in their response to inflammatory mediators is not understood. To study permeability regulatory processes on an individual cell basis in intact microvessels, we developed methods to delineate individual endothelial cells within living, single perfused microvessels.</jats:p><jats:p><jats:bold>Methods:</jats:bold> Individual postcapillary microvessels in the mesenteries of frogs and hamsters were perfused with chloride‐free Ringer's solutions containing AgNO<jats:sub>3</jats:sub> (0.1 g per 100 ml) for 5–10 s. Vessels were immediately flushed with Ringer's solution containing serum albumin and some vessels subsequently perfused with the fluorescent nucleic acid stain YO‐PR0‐1 (1 μM) for 10–15 min. Vessels were imaged in situ using laser scanning confocal microscopy. A reflectance image from silver precipitate in the endothelial clefts and a fluorescence image of the nuclei were simultaneously recorded. Stacks of confocal images were merged and used to reconstruct the three‐dimensional orientation of endothelial cells. Hydraulic conductivity of some frog mesenteric venular vessels, measured before and after perfusion with AgNO<jats:sub>3</jats:sub>, was used to assess the integrity of the vessels.</jats:p><jats:p><jats:bold>Results:</jats:bold> The endothelial cell clefts were delineated with a very fine, readily imaged precipitate of silver. Hydraulic conductivity measured after AgNO<jats:sub>3</jats:sub> perfusion in frog mesenteric venular microvessels was not significantly different from control. Vessels showed a slight reduction in reactivity to inflammatory stimuli. The three‐dimensional pattern of endothelial cells in the living vessels was imaged.</jats:p><jats:p><jats:bold>Conclusions:</jats:bold> Combining confocal microscopy with silver staining in single perfused microvessels was shown to be an effective means to delineate the three‐dimensional pattern of endothelial cells forming microvessel walls. It enables further study of the vascular structure and function relationships at the individual cell level in intact microvessels.</jats:p> Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy Microcirculation |
| spellingShingle | He, P., Adamson, R. H., Microcirculation, Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy, Physiology (medical), Cardiology and Cardiovascular Medicine, Molecular Biology, Physiology |
| title | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_full | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_fullStr | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_full_unstemmed | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_short | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| title_sort | visualization of endothelial clefts and nuclei in living microvessels with combined reflectance and fluorescence confocal microscopy |
| title_unstemmed | Visualization of Endothelial Clefts and Nuclei in Living Microvessels with Combined Reflectance and Fluorescence Confocal Microscopy |
| topic | Physiology (medical), Cardiology and Cardiovascular Medicine, Molecular Biology, Physiology |
| url | http://dx.doi.org/10.3109/10739689509146772 |