We're getting reasonably good results using standard Karnofsky's fixation, OsO4, uranyl acetate & lead citrate staining, but I've heard of people using other stains - particularly ruthenium red - to stain the proteoglycans in the ECM better.
I'd also be interested if anyone has found good EMs of fish ECM in the literature.
Cheers
Hi,
dear Bryan, there are other possibilities (in addition to the classical aldehyde 'Karnovsky' fixative, UO2Ac - PbCitrate section staining .....) unfortunately you say nothing about the PG-type you are after...(or do you expect only "excellent imaging of 'PG's' generally? Unfortunately I haven't done research on fish ECM (which perhaps - compared to human tissue might differ regarding composition and chemistry/physics of ECM-components). Usually OsO4-treatment would be enough, but RuO4 perhaps will precipitate more molecules.....(thus leaving the "natural hydrated space" of PG-molecules more/less "empty")
For being short I only mention substances for now (applied/added to fixative or even as as staining component):
CPC, (OsO4-, RuO2, RuO4, RuRed & other chemicals)-PPD (mordanting)-treatment, Lanthanum(nitrate)/lanthanides, ....
e. g. cf: https://www.researchgate.net/publication/200473363_Three-dimensional%27tubular-like%27_structures_in_the_matrix_of_aged_human_costal_cartilage_artefact_or_morphological_equivalent_of_Proteoglycan-aggregates?
Article Three-dimensional,'tubular-like' structures in the matrix of...
Check the new imaging technique that eliminates the use of stains for optically anisotropic biological entities. Using this technique you can look at average molecular orientation of your embryos and the detailed structure.
Check the device at www.polaviz.com
www.polaviz.com
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