Animal Dissection - Science topic

You can injeact the spleen with warm phosphate buffer saline and incubate the spleen for 30 minutes at 37, then mix the spleen content well then withdraw the content.

Hi Eric, I am not really sure what you meen under "microdissected speific regions".

Just in case you have cut the brain conronally or horizontally on a cryostate or vibratom then you have the possibility to mount, let me say, evera 6th section on a slide. After air drying you can stain  the sections with cresylviolett. This so called Nissl-stain helps orientating.  If you need the whole block where your structure of interest is in,  you have to use the coordinates of the Pxinos-Brain-Atlas. Good luck!

You can check these articles

Dear Janine,

To maintain colocalization try embedding the bone marrow in a gelatin solution, 8-10%, prior to freezing, this should keep the cells intact for sectioning.

Hi Malavika, a smaller needle will surely get your cells disrupted better and faster, if and only if the pore is not clogged. So it depends on the type of cells you are homogenizing and also which part of the cells contain the enzyme you are assaying.

Many thanks Abdelaziz 

Thank you Tiago 

really, this article is useful

We have published a research paper entitled "Parasympathomimetic Effect of Shilajit Accounts for Relaxation of Rat's corpus cavernosum"; reference: Am J Mens Health March 2013 vol. 7 no. 2 119-127. This paper may be of use for you. 

Hi Jasmine,I apologize for not replying earlier.

I have tried many ways for uncoiling the Fallopian tube .However,after many trials I decided to not uncoil the oviduct, because it misplaces the sperm. Instead, I gently dissect out the fallopian tube  and place it in about 50 ul medium between two cover-slips supported by silicon grease. In that way, I can flip the oviduct over and back in order to get the best views of the sperm in various coils. hope that's somewhat helpful.

Best,

Liliya

I do not know that whether Magnesium Sulfate can be detected on postmortem examination.  but I know that Magnesium Sulfate make all never and muscle relaxed, painless and easy to death (euthanasia)....... However, you may compare with and without the higher Magnesium Sulfate' postmortem muscle tissue sections to see whether there are different, or look for the method which can be detected the concentration of Magnesium Sulfate on the postmortem muscle tissues......

Sorry Ana... I had forgotton to thank you. But I had watched the video already.

Hi Jeff,

I have done the extraction on rats  ( 250 gm weight) and it is was smooth, I imagine the same would apply to mice.

what I did is prepare an ice cold sterile(autoclaved) normal saline, a 60 ml syringe and an 11 gauge needle( blunted tip) now after deep anesthesia the rat spinal column with all surrounding soft tissue is removed from the 3 cervical to the lower L5 segment.

the syringe is filled up and the needle is mounted on the syringe. the lower ( lumbar part is searched for the vertebral foramen and when the opening is identified, the tip pf the needle is inserted in the spinal canal. ( 1cm deep) make sure that the needle and the spinal column are alligned to same plane ( together creating a perfect straight line and,  it is tight enough to prevent back leakage and a forceful push( in one movement, the plunger of the syringe is pushed ( emphasis on one shot push , not advancing the plunger gradually) and the spinal cord is flushed out at the cervical end in one piece intact ( flushed into sterile stainless steel kidney shape tray with sterile normal saline) the cauda equina is identified and the alignment of the spinal cord is done accordingly ( rostral end - and caudal end .

in mouse case, I imagine the needle gauge should be a tight fit to the LS vertebral foramen, and the syringe ( 10 ml or 15 ml should suffice, but to make sure you need to try to remove the spinal column with all soft tissue and match till you find a suitable needle gauge size to tightly fit to L5, and make several trials till it works .

best wishes

A few possibilities exist:

1) You could rinse the tissues or go through a perfusion protocol with the organs before sectioning them.

2) You could potentially treat the tissues with a peroxidase inhibitor, lowering the numbers of RBCs present in the tissue.

3) Finally, I found a protocol for lysing RBCs in a tissue sample (see attached pdf).

Let me know if any of these work out for you. If not, I'm happy to continue looking.

Hello

I think it would be more about the depolarisation of the neurons (due to the static charges on the metal object), which would lead to preventing any transmission. Of course this is a problem only, when such properties are required for the research.

For some experiments you may be able to model processes in immortalized cell lines which are no longer maintained by animal sacrifice. From a research perspective it'll depend on your hypothesis. Financially it may also be cheaper to use live animals unfortunately. 

Another alternative is to try and get more "mileage" out of each animal using repeated experimentation, survival surgery, or fixed/frozen tissue archives.

The following paper will give you detailed instructions for the dissection of different brain areas, including striatum (caudate putamen and globus pallidus) and hippocampus:

Glowinski J, Iversen LL.

Regional studies of catecholamines in the rat brain. I. The disposition of [3H]norepinephrine, [3H]dopamine and [3H]dopa in various regions of the brain.

J Neurochem. 1966 Aug;13(8):655-69

Well it is a description for the dissection of various rat brain areas, but I could use this instruction also for mouse brains (mouse brains will need more preparation skills because its approx. 4 to 5 times smaller than rat brain, but anatomically they are very similar).

If you need further assistance please contact me via email, I think I have also step by step photos of this dissection procedure.

Best,

Murat

Hi Lakshmi Narendra,

You could refer to a book by Palkovits on brain maps. The title is : Maps and guide to micro-dissection of the rat brain.

If you would prefer videos as suggested by Ramirez-Franco,  Jove is a good place to look. Take a look at these as well:

Hope this helps!

Take the slice from which you dissect your regions of interest, fix the remaining tissue and slice using a microtome/cryostat. Then stain the section using e.g. cresyl violet and validate based on anatomy

I would recommend you to remove the two inguinal lymph nodes: they are big and easy to find (see attachment: look at the intersection of the 3 veins). Simply pull it out using forceps

Thanks! It is what I am looking for!