Additionally, I find varicosities in some neurons. I am puzzled as to what they amount to?
What do you mean by "Dead neurons"? Your block of tissue first should be fixed up using appropriate fixatives, then follow the Golgi techniques: Rio-Ortega is one of the best. About varicosities; depends on animals and your experimental design, usually the bulging appears at the exits of primary dendrites from soma.
Formalin-fixed brains can be impregnated by the Golgi technique. Check the papers for techniqual detailes published in the 1980th by Seress L and Ribak C.
In Golgi techniques,varicosities are located on axons and spines are visible on dendrites.
Dendrites with varicosities along their length are sometimes referred to as moniliform. Electon microscopic examination shows that these varicosities are filled with mitochondria.
Thank you all,
Gila, that was exactly what i observed,
Ilsa, your point seems interesting, can you please let me know the author of that particular paper
Thank you all once again
Hi
I have used a single section Golgi technique to label scrapie infected neurons this technique is much quicker than the block technique. I did move on tosingle Lucifer yellow injections of neurons in order to quantify using the confocal microscope. Can send you relevant publications if you wish ??
Debbie
Dear Debbie, I am interested to know about too, it would be grateful if you send me the relevant publication if you don't mind. Thanking you in advance,
Gila
According to the Principles of Neuropsychology book by Eric Zillmer (2007) the silver chromate stain dead neurons in black:
https://books.google.de/books?id=XYoFAAAAQBAJ&pg=PA34&lpg=PA34&dq=golgi+stain+dead+neurons&source=bl&ots=SPPHbGjUac&sig=WUJQ1ZrBC87QsYsw1AxYsbbnIMA&hl=es&sa=X&ei=1zipVPPXGqS9ygOb_4FY&ved=0CDgQ6AEwBA#v=onepage&q=golgi%20stain%20dead%20neurons&f=false
I have limited experience with Golgi staining, but have found that glial cells, blood vessel epithelial cells and a variety of other things occasionally take up the stain. I would guess that not ALL the neurons stained are dead or dying, but it would be difficult to test this.
The animals should be transcardially perfused with 4 % cold formaldehide (4-6 degree C). The next day the brain or spinal cord should be removed. Cut 5 mm thick tissue blocks and place them into a solution containing 4 % potassium dichromate and 1 % osmium tetroxide in distilled water. This solution should be kept in dark tightly closed glass bottles (osmium is dangerous and volatile!). Place 3-6 tissue blocks, separated from each other to 50 ml of solution. After 3-4 days transfer the blocks into 0,75 % silver nitrate solutions. A redish-brown presipitate will be formed. Rinsed the blocks in this silver solution, until it remains clear, then store the bloks in this solution for 3 days. Blocks should be embedded in Celloidine or some similar medium. Cut 60-100 micro meter thick sections. The Golgi technique is difficult, the result is varibale. I used this technique to impregnate cerebellum, cortex, spinal cord of young 3-4 weeks old kittens, rabbits, rats for teaching purposes. Myelinated axons are usually not impregnated. Formation of precipitate is frequent.
I used Golgi to study the development os Xenopus laevis CNS. You can see my paper in Research Gate under my name. (Development of the optic tectum in Xenopus laevis, J. Anatomy, 116/3, 347-353, 1974)
Good luck!
Hi Yogesh Kanna Sathyamoorthy ,
I'm currently analyzing dendrites after golgi-cox staining and noticing varicosities on my dendrites as well. Did you happen to figure out the reason for this?
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