I have several deep-sea bivalve specimens, and I have observed that the staining efficiency of Mutvei is not satisfactory in the shell sections. Only a small portion of the growth lines appear clearly stained. Could someone provide me with potential factors that may be influencing this issue or suggest some improvement measures
Dear Zhang Xiaochun would you mind to tell people in this forum (which may be inexperienced or even naiv with regard to that really interesting technique to examine e.g. accretionary hard parts of organisms like shells or skeletal structures of other calcified marine detritus consisting of biogenic carbonates and calcium phosphates) which specific literature you may have read already and - much more important - which 'recipe' for the 'Mutvei's solution' you actually are following, how you prepare (specimen prep) your deep-sea bivalve shells (I 'guess': will be embedded into resin (?, which one...), then getting - by a diamond saw - 2-3 mm thick sections, then ground to >thin sections< and then etched, fixed and stained by the mentioned and used 'Mutvei's Solution'.
"Potential factors" of failure (or - at least - poor outcome) of prominent staining AND/or STRUCTURE linings almost are uncountable....especially if one doesn't know in detail what has been done with the specimens.
I myself have to admit that I am not "practically" familiar with staining of 'cutting-grinding techniques' and haven't either prepared shells or calcified material or used the Method per se, but I am familiar with the 'ingredients' of the Mutvei's solution as well as most of the preparation steps (e.g. embedding, and treatment of thick/thin sections ending up finally with the etching, fixing and staining steps to be done.
NB: edit 23-11-18: deap-sea ==> deep-sea (this correction only)
Dear Wolfgang H. Muss,Thank you for your attention to the question! The sample preparation is similar to what you initially hypothesized. I sliced the sample approximately 1cm thick along the maximum growth axis of the shell, and then carried out grinding and polishing. Subsequently, I employed the classic staining protocol (Schöne et al., 2005), which involved etching at 38°C for 10-20 minutes. Overall, the sample processing procedure is comparable to those reported in previous studies.
However, the distinct characteristic of my shell samples is their high curvature (resembling a banana shape). During the cutting process, I had to avoid heavily worn areas on the shell's apex, which may have resulted in suboptimal visualization of the growth lines. Nevertheless, it is also possible that this issue is due to the unique nature of deep-sea samples, as all my specimens exhibited a similar problem. In addition, I tried to extend the staining time, but the results were not significant, and even blotchy, bubble-like things appeared on the sections.
Most of the shells displayed clear growth lines in the middle region of the outer shell, while other areas were either not visible due to wear or had indistinct growth lines that were difficult to observe. It is currently unclear if the discontinuous and blurry regions are a result of staining issues or other factors. I am currently improving my experimental procedure to gradually eliminate any unfavorable influences.
Dear Zhang Xiaochun , since I found again this your interesting and informative answer - +/- 'accidentally - just some minutes ago:
Thank you so much for elucidating some details of your specimen processing, the problems you encounter. and the method of staining you use (SCHÖNE = (following the English notation style:) SCHOENE et al, 2005; cf.: https://www.sciencedirect.com/science/article/abs/pii/S0031018205002804 ). It seems that the article/paper isn't in Research Gate's archive of publications, otherwise it would pop off here with the title
'Mutvei's solution: An ideal agent for resolving microgrowth structures of biogenic carbonates'
by Bernd R. Schöne, Elena Dunca, Jens Fiebig, Miriam Pfeiffer' within the 'green marked hyperlink.....
ah... see now that on entering the original DOI-address (https__://doi.org/10.1016/j.palaeo.2005.03.054, vide the 2 underscore signs which have to be deleted in a/the browser before executing) 'mutates' the original LINK to
[Article Mutvei's solution: An ideal agent for resolving microgrowth ...
]For all those not familiar with that kind of etching-staining solution, I'd like to mention - at least - its composition here (also to aid in further keyword-search):
>One liter of Mutvei's solution consists of 500 ml 1% acetic acid, 500 ml 25% glutaraldehyde and ca. 5 to 10 g alcian blue powder
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