Two-color fluorescence labeling in acrolein-fixed brain tissue
SUMMARY Acrolein is a potent fixative that provides both excellent preservation of ultrastructural morphology and retention of antigenicity, thus it is frequently used for immunocytochemical detection of antigens at the electron microscopic level. However, acrolein is not commonly used for fluore...
| Main Authors: | , , , |
|---|---|
| Format: | info:eu-repo/semantics/article |
| Language: | eng |
| Published: |
Histochemical Society
2010
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10171/13044 |
| _version_ | 1793400051106578432 |
|---|---|
| author | Luquin, E. (Esther) Lorenzo, E. Aymerich-Soler, M.S. (María Soledad) Mengual, E. (Elisa) |
| author_facet | Luquin, E. (Esther) Lorenzo, E. Aymerich-Soler, M.S. (María Soledad) Mengual, E. (Elisa) |
| author_sort | Luquin, E. (Esther) |
| collection | DSpace |
| description | SUMMARY Acrolein is a potent fixative that provides both excellent preservation of ultrastructural
morphology and retention of antigenicity, thus it is frequently used for immunocytochemical
detection of antigens at the electron microscopic level. However, acrolein is not
commonly used for fluorescence microscopy because of concerns about possible autofluorescence
and destruction of the luminosity of fluorescent dyes. Here we describe a simple protocol
that allows fine visualization of two fluorescent markers in 40-mm sections from acroleinperfused
rat brain. Autofluorescence was removed by pretreatment with 1% sodium borohydride
for 30 min, and subsequent incubation in a 50% ethanol solution containing 0.3%
hydrogen peroxide enhanced fluorescence labeling. Thus, fluorescence labeling can be used
for high-quality detection of markers in tissue perfused with acrolein. Furthermore, adjacent
acrolein-fixed sections from a single experiment can be processed to produce high-quality
results for electron microscopy or fluorescence labeling. |
| format | info:eu-repo/semantics/article |
| id | oai:dadun.unav.edu:10171-13044 |
| institution | Universidad de Navarra |
| language | eng |
| publishDate | 2010 |
| publisher | Histochemical Society |
| record_format | dspace |
| spelling | oai:dadun.unav.edu:10171-130442024-01-30T13:56:29Z Two-color fluorescence labeling in acrolein-fixed brain tissue Luquin, E. (Esther) Lorenzo, E. Aymerich-Soler, M.S. (María Soledad) Mengual, E. (Elisa) sodium borohydride endogenous peroxidase inactivation autofluorescence ethanol enhanced penetration hydrogen peroxide aldehyde fixation SUMMARY Acrolein is a potent fixative that provides both excellent preservation of ultrastructural morphology and retention of antigenicity, thus it is frequently used for immunocytochemical detection of antigens at the electron microscopic level. However, acrolein is not commonly used for fluorescence microscopy because of concerns about possible autofluorescence and destruction of the luminosity of fluorescent dyes. Here we describe a simple protocol that allows fine visualization of two fluorescent markers in 40-mm sections from acroleinperfused rat brain. Autofluorescence was removed by pretreatment with 1% sodium borohydride for 30 min, and subsequent incubation in a 50% ethanol solution containing 0.3% hydrogen peroxide enhanced fluorescence labeling. Thus, fluorescence labeling can be used for high-quality detection of markers in tissue perfused with acrolein. Furthermore, adjacent acrolein-fixed sections from a single experiment can be processed to produce high-quality results for electron microscopy or fluorescence labeling. 2010-09-27T11:30:40Z 2010-09-27T11:30:40Z 2009-12-16 info:eu-repo/semantics/article https://hdl.handle.net/10171/13044 eng info:eu-repo/semantics/closedAccess Histochemical Society |
| spellingShingle | sodium borohydride endogenous peroxidase inactivation autofluorescence ethanol enhanced penetration hydrogen peroxide aldehyde fixation Luquin, E. (Esther) Lorenzo, E. Aymerich-Soler, M.S. (María Soledad) Mengual, E. (Elisa) Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title | Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title_full | Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title_fullStr | Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title_full_unstemmed | Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title_short | Two-color fluorescence labeling in acrolein-fixed brain tissue |
| title_sort | two-color fluorescence labeling in acrolein-fixed brain tissue |
| topic | sodium borohydride endogenous peroxidase inactivation autofluorescence ethanol enhanced penetration hydrogen peroxide aldehyde fixation |
| url | https://hdl.handle.net/10171/13044 |
| work_keys_str_mv | AT luquineesther twocolorfluorescencelabelinginacroleinfixedbraintissue AT lorenzoe twocolorfluorescencelabelinginacroleinfixedbraintissue AT aymerichsolermsmariasoledad twocolorfluorescencelabelinginacroleinfixedbraintissue AT mengualeelisa twocolorfluorescencelabelinginacroleinfixedbraintissue |