Hi everyone,
I obtained TEM images from suspended MCF10A mammary epithelial cells treated with a chemical.
Since I'm not familiar with interpreting TEM images, I want to get a valuable advice from all of you.
I annotated some parts as red arrows in the image I uploaded.
Is it ER swelling or other cellular changes?
Would you please give me your interpretation?
Thanks a lot!
Hello,
My first thought is plasma membrane changes. The white is at the exterior of the cell.
I'd agree with Nicolas Barois. Looks like folding of the plasma membrane to me. You can make serial sections of this sample to confirm this.
The rER visible here looks normal.
Those are tightly packed microvilli in lateral intercellular spaces. In this paper they are more spread out. https://www.researchgate.net/publication/307593144_Involvement_of_Rho_GAP_GRAF1_in_maintenance_of_epithelial_phenotype
Jeongsik Kim :
Seconding opinions of Nicolas Barois and Martin Schauflinger .
Under the conditions your MCF-10A cells have been "treated with 'a chemical' and then have been processed unto EM-TEM-visualization you /one might expect 'unphysiological or unspecific' cellular ultrastructural changes and / or behaviour,
(e.g. by cellular stress, due to culture conditions and treatment, and - last but not least: your Specimen processing conditions) as well as 'experimentally induced "true" alterations / changes of cellular compartments - since the cells were treated by the / a chemical.
Therefore it would be helpful or of benefit to get some details of the culture conditions as well as (at least - 'major' - ) specimen processing details (chemically fixed ?/ HPF? etc.) to be able to perhaps evaluate real "treatment"-induced cellular alterations.
(BTW: congratulations regarding the achieved quality of the illustrative EM-image...)
What about reading / take into consideration:
UC Berkeley
UC Berkeley Electronic Theses and Dissertations
Title: Ultrastructural Characterization of Cellular Interactions Within Resting, Developing and Tumorigenic Breast Epithelium.
Look for the Specimen Processing (Experimental Procedures, p.47 ff.), especially
descriptions for microvilli / Filopodia (=actin rich membrane proteins), 'microvilli-like structures/ inter-cellular membrane protrusions / cytoplasmic membrane proliferations (are not filopodia)...; >>Invadopodia (and podosomes );
(quote from p.74:) "....The importance of the proper physiologically relevant 3D environment is supported by recent evidence that demonstrates differences in cellmigration of fibroblasts grown in cell derived matrix and pure collagen 1 matrix ..."(end quote, )
cf. text p.14; fig. 7, p.22; '...prevalent membrane protrusions along the interior facing surfaces of the cells....'(p. 42) - images & legend Fig. 16 (p.41) and text p.44 - 45,
Permalinkhttps://escholarship.org/uc/item/2bt0f430
Author: JORGENS, Danielle Mary
Publication Date: 2013
Peer reviewed | Thesis / dissertation
https://escholarship.org/content/qt2bt0f430/qt2bt0f430.pdf
WANG et al, 2013:
PNAS January 2, 2013 110 (1) 163-168; first published December 17, 2012; https://doi.org/10.1073/pnas.1201141110
'Rotational motion during three-dimensional morphogenesis of mammary epithelial acini relates to laminin matrix assembly.'
https://www.pnas.org/content/110/1/163
TANG et al, 2014:
Report: Plasticity of Mammary Cell Boundaries Governed by EGFand Actin Remodeling
Cell Reports 8, 1722–1730 September 25, 2014 ª2014 The Authors http://dx.doi.org/10.1016/j.celrep.2014.08.026
Supplemental Information includes Supplemental Experimental Procedures,
five figures, and five movies and can be found with this article online at
http://dx.doi.org/10.1016/j.celrep.2014.08.026.
Hello Jeongsik Kim,
If you want a clear and fast picture on the effect of your treatment, you should do SEM on cell monolayer on glass coverslips, treated and untreated.
In TEM, you can also do transversal sectioning of the cell monolayers, grown on transwell filter or on glass coverslip (in this case flat embedding on the glass and removing it with hydrofluoric acid - harmful).
As for me, it looks like formation of lytic vacuoles. Like that one in the upper part of the image, red arrow (the left one) crosses it.
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