I have perfused and frozen rat brain tissue (from a new transgenic line) that I have cryosectioned onto slides in order to characterise a number of different cell types. One thing I would like to check is whether these animals have a difference in the density and morphology of spines on their neurons.
My initial research suggests that the golgi stain would require a different preparation of the tissue, and as I am hoping to do a lot of different stains with one tissue I would prefer not to sacrifice a whole brain for this stain if this is not something worth pursuing!
Is there a way I can look at spines on neurons using the tissue I have already?
Dear Sophie Quick :
"Perfused" = means already "fixed brain tissue"? if so, please define the fixative you used and at least the more important prep-steps until freezing your specs and cryo-preservation.
Ment only as reading proposals:
ZHANG et al, 2020
Front Neuroanat. 2020; 14: 585513.
(= https://www.frontiersin.org/articles/10.3389/fnana.2020.585513/full and PDF)
Published online 2020 Nov 9. doi: 10.3389/fnana.2020.585513
>Optimized Golgi-Cox Staining Validated in the Hippocampus of Spared Nerve Injury Mouse ModelStep by Step Golgi-Cox Staining for Cryosection<
PROTOCOLS Article
Front. Neuroanat., 19 June 2019 DOI: Download full text also in:Article Step by Step Golgi-Cox Staining for Cryosection
|[Unfortunately cryoprocessing/freezing only, no initial chemical fixation given]
For Open Access: cf.: https://www.frontiersin.org/articles/10.3389/fnana.2019.00062/full
DU 2019:
doi: 10.1002/cpns.69.. 2019 Jun;88(1):e69.
Curr Protoc Neurosci
>Golgi-Cox Staining of Neuronal Dendrites and Dendritic Spines With FD Rapid GolgiStain™ Kit< cf.: https://pubmed.ncbi.nlm.nih.gov/31216393/ or
https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpns.69 [Article pdf for non-subscribers available only as PPV = Pay-per-view).
CHENG et al, 2014:
Methods Article
>Fluorescent labeling of dendritic spines in cell cultures with the carbocyanine dye “DiI”Nissl Staining Method and Protocol on Frozen or Vibratome Sections for Brain & Spinal CordImmunohistochemical labelling of spinal cord neurons involved in bladder activity< i.e. (quote): "....Cryoprotect fixed tissue (L5-S2 spinal cord)..." (end quote)
https://www.protocols.io/view/immunohistochemical-labelling-of-spinal-cord-neuro-bakkicuw
Ke NING (et al; University of Sheffield, Sheffield, UK), TREAT NMD -Neuromuscular Network: SMA_M.1.2.004;
Issued September 17th 2010 Last reviewed July 11th, 2017
SOP: "Cell Body-Motor Neuron Counts" (see Methods section)
http://www.treat-nmd.eu/downloads/file/sops/sma/SMA_M.1.2.004.pdf
More to discover on "deep digging Google and / or specific literature...
Best of luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue