I was trying to get an easier/improved method for Thioflavin S staining and came across different authors some of which said Thioflavin S stains tau-based neurofibrillary tangles/paired helical filaments (NFTs/PHF) (Sun, Nguyen, and Bing 2002; Liu et al. 2012; Santa-Maria et al. 2007)as against the Amyloid βeta plaques that I’ve always thought Thioflavin S labels (Bussière et al. 2004; Ly, Cai, and Song 2011; Urbanc et al. 2002).
Can someone kindly help clarify this? Given that both Aβ -plaques and NFTs are expected to be found in Alzheimer’s disease brains, how then does one interpret ThioflavinS-positive inclusions? Do you just say it’s both or you choose whichever one you prefer? Any help will be appreciated.
The references are here attached.
REFERENCES
Bussière, Thierry, Frédérique Bard, Robin Barbour, Henry Grajeda, Terry Guido, Karen Khan, Dale Schenk, Dora Games, Peter Seubert, and Manuel Buttini. 2004. “Morphological Characterization of Thioflavin-S-Positive Amyloid Plaques in Transgenic Alzheimer Mice and Effect of Passive Aβ Immunotherapy on Their Clearance.” The American Journal of Pathology 165 (3) (September): 987–995.
Liu, Li, Valerie Drouet, Jessica W. Wu, Menno P. Witter, Scott A. Small, Catherine Clelland, and Karen Duff. 2012. “Trans-Synaptic Spread of Tau Pathology In Vivo.” PLoS ONE 7 (2) (February 1): e31302. doi:10.1371/journal.pone.0031302.
Ly, Philip T.T., Fang Cai, and Weihong Song. 2011. “Detection of Neuritic Plaques in Alzheimer’s Disease Mouse Model.” Journal of Visualized Experiments (53) (July 26). doi:10.3791/2831. http://www.jove.com/video/2831/detection-of-neuritic-plaques-in-alzheimer-s-disease-mouse-model.
Santa-Maria, Ismael, Félix Hernández, Joaquín Del Rio, Francisco J. Moreno, and Jesús Avila. 2007. “Tramiprosate, a Drug of Potential Interest for the Treatment of Alzheimer’s Disease, Promotes an Abnormal Aggregation of Tau.” Molecular Neurodegeneration 2 (1) (September 6): 17. doi:10.1186/1750-1326-2-17.
Sun, Anyang, Xuan V. Nguyen, and Guoying Bing. 2002. “Comparative Analysis of an Improved Thioflavin-S Stain, Gallyas Silver Stain, and Immunohistochemistry for Neurofibrillary Tangle Demonstration on the Same Sections.” Journal of Histochemistry & Cytochemistry 50 (4) (April 1): 463–472. doi:10.1177/002215540205000403.
Urbanc, B., L. Cruz, R. Le, J. Sanders, K. Hsiao Ashe, K. Duff, H. E. Stanley, M. C. Irizarry, and B. T. Hyman. 2002. “Neurotoxic Effects of Thioflavin S-positive Amyloid Deposits in Transgenic Mice and Alzheimer’s Disease.” Proceedings of the National Academy of Sciences 99 (22) (October 29): 13990–13995. doi:10.1073/pnas.222433299.
Thioflavin S labels beta-pleated sheets, so it would include plaques, tangles, neuropil threads, and vascular amyloid. NFTs and plaques labeled this way look different, and threads are easily distinguishable.
As Patrick indicated thio-s labels beta pleated sheets. The neurofibrillary tangles will be mature and extracellular (ghost) tangles. This is quite helpful for interpretation of neuronal loss. You will be able to see an assortment of plaques (diffuse/primitive, compact, cored, and neuritic). Additionally, amyloid angiopathy will be visible. Lewy bodies are not as easily seen using thio-s.
ThioS labels both. You can distinguish tau aggregates with fluorescence microscopy based on their shape: NFTs are distinctively flame shaped and are smaller than beta-amyloid plaques which are large round structures with or without highly fluorescent core.
Thank you all. I should have thought about site (with respect to the cell) and shape; i was. Thanks once again for your very helpful responses.
Thiofavin S stains both, Amylod plaques and Tangles. Here are a few references for the protocol (see attached links)
http://www.alzforum.org/res/com/protocol/tmp/2009022591c57f6c5f2d4e4391a3282b42a03439.pdf; http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2990.1986.tb00677.x/abstract
Tau specific stains exist. Steingrimur Steffanson and others have put together a list of fluorescent probes for amyloid at https://www.researchgate.net/project/Amyloid_open_protocols/
Message me if you want to join.
I have provided an image of the assortment of Alzheimer's disease inclusions, including diffuse plaques which are visible.
Thanks Melisssa; these pictures are so very helpful. Can you kindly provide a reference for the image? Thank you
@Melissa: Thanks a lot for your image. I would like to ask one silly question here. How do you actually differentiate between diffuse and neurite plaques or specifically how one can define neuritic plaque.
Secondly, I recently performed Thioflavin-S staining procedure to visualise amyloid plaques in the tissue sections of Alzheimer Disease mouse brain. I'm presenting an image of Thioflavin- S stained amyloid plaques in the cortical region.
Hence, I am looking for proper methods to quantitate the amyloid plaques in my images which I can use for further analysis with respect to percent plaques load per area. Can you please guide me as how to proceed and what all plaques to choose for the same. Please help me in as to how can I go about quantitating the amyloid plaques and which software/tools to use?
@Samuel. I put together the figure for you yesterday, so it does not have a reference. I uploaded a manuscript to my Research Gate page from our atypical AD study. It does have a THioflavin-S image of neurofibrillary tangles, dense core plaque, and a neuritic plaque. Please let me know if this is sufficient.
https://www.researchgate.net/publication/51533664_Neuropathologically_defined_subtypes_of_Alzheimer's_disease_with_distinct_clinical_characteristics_a_retrospective_study
Article Neuropathologically defined subtypes of Alzheimer's disease ...
@Apurva. I replied to your message, let me know if it went through okay. I also attached a figure that might help.
There is another thread that I recommend for the quantification of amyloid burden.
https://www.researchgate.net/post/Best_Image_analysis_software_to_quantify_IHC_markers_in_tissue_microarrays
Non-Neuritic and Neuritic plaque (double stained with amyloid and tau). It is the tau pathology that defines the neuritic plaque, not the dystrophic (kind of twisted) nature of the the neurites.
@Melissa: I am sorry but I did not receive the message. Can you please send it again...
good pictures from Melissa- but just to add a small note- Thio-S will not label "pre-tangles". Thio-S can also label Lewy bodies which are found in roughly 50% of AD cases (but they are not as bright as plaques and tangles).
If you are on a budget (although always use at least two stains to confirm any deposits) Thio-S is very cheap and quick- as compared to Silver stains or IHC which can drive up your cost 10 fold.
@Melissa @Apruva Agrawal sorry it has been awhile, but would either of you happen to have a thioflavin S quantification protocol for image J? Thanks for your help!
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