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Influence of chemical and freezing fixation methods in the freeze-fracture of stratum corneum
- Source :
- Digital.CSIC. Repositorio Institucional del CSIC, instname
- Publication Year :
- 2003
-
Abstract
- 8 pages, 7 figures.-- PMID: 15099572 [PubMed].-- Printed version published Jun, 2004.-- Corrigendum published in: Journal of Structural Biology 148(2): 257 (Nov 2004), http://dx.doi.org/10.1016/j.jsb.2004.06.001<br />A comparison between two fixation techniques for freeze-fracture was established. Stratum corneum (SC) samples from pig epidermis were fixed using high-pressure freezing (HPF) and using plunging in propane freezing; the latter after chemical fixation. Then, frozen samples were freeze-fractured, coated with platinum–carbon, and visualized using a high-resolution low-temperature scanning electron microscope and a transmission electron microscope. Our results indicate that the plane of freeze-fracture was different depending on the fixation and freezing methodology used. In the samples frozen by HPF without chemical fixation, the fracture plane laid mainly between the lipid lamellae. However, when chemical fixation and plunging in propane freezing was used, the fracture plane did not show preference to a specific way. Plunging in propane freezing of chemically fixed samples, on the other hand, provides a more homogeneous fracture behaviour. Thus, depending on the methodology used, we can favour a visualization of either lipid or protein domains of the SC. These results could be very useful in future ultrastructural studies in order to facilitate the microscopic visualization and interpretation of the complex images such as those of SC and even of other samples in which different domains coexist.<br />This study was supported mainly by funds from DGICYT (Dirección General de Investigación científica y técnica) (Prog. No. 2001-1188-CO2-02).
- Subjects :
- Tissue Fixation
Scanning electron microscope
Chemical fixation
Swine
Analytical chemistry
High-resolution low-temperature SEM
Stratum corneum
High-pressure freezing fixation
Propane
Structural Biology
medicine
Animals
Freeze Fracturing
Composite material
Fixation (histology)
Chemistry
Cryoelectron Microscopy
Freeze-fracture
Proteins
Fixation method
Lipids
medicine.anatomical_structure
Homogeneous
Transmission electron microscopy
Ultrastructure
Fracture (geology)
Indicators and Reagents
Epidermis
Subjects
Details
- ISSN :
- 10478477
- Volume :
- 146
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Journal of structural biology
- Accession number :
- edsair.doi.dedup.....85515544dc08766d764506310dd2014d