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I was trying to isolate fibroblastic reticular cells (FRC) and follicular dendritic cells from mice spleen, but kept getting poor yield. I found from literature that the frequency of FDC is around 0.2% in spleen but I could not find data about FRC. What is the expected frequency of FRC in mice spleen? Thanks.
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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.
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I have whole mouse brains fixed in PFA that I need to microdissect specific regions: hippocampus, cerebellum, striatum,and frontal cortex for DNA methylation analysis. Is there a staining (or any other) technique I could use to verify that I have dissected the region I intended, and that there is not contamination with surrounding regions? 
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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!
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HI 
Actually, I want to know the possible way and procedure to sample CSF from cisterna magna of Rat brain without any surgery and opening the skin in this area. 
thanks
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I would like to perform immunofluorecence on human bone marrow biopsies which are snap frozen. I received bone marrow biopsies from the tibia (spongy material, 1x1 cm) which I embedded in OCT and froze in 2-methyl-butane. Unfortunately, cutting sections of this material seems to be impossible since the tissue ruptures very easily. Does someone know how to optimize this protocol and prepare the bone marrow sections?
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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.
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I know that the needle size will affect disruption of the cells, but do cells still get disrupted at the larger size? TIA
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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.
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I am trying to isolate total RNA from tadpoles for transcriptomics studies. I have to do for the first time so if someone has a protocol.please share with me
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Many thanks Abdelaziz 
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Hey. For osteology fish. Someone has a protocol or an approach to dissect and visualize the bone head?
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Thank you Tiago 
really, this article is useful
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I want to dissect corpus cavernosal tissue for organ bath studies in rat. Please send me links where from i can learn them to dissect.
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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. 
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I am trying to track sperm cells in the oviduct. However, I am facing some difficulties uncoiling the oviduct.I am preforming the dissection on a super-ovulated female a few hours after mating and I cannot seem to locate where to make the incision/the cut in order to remove the ovarian bursa without harming the oviduct itself.
Thank you
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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
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Does Magnesium sulfate  used for euthanasia produce any visible lesions that can be detected on postmortem examination?
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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......
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I am working on a teleost fish from Cyprinidae family. I have read many procedures for head kidney (HK) isolation and maintaining its culture but if anyone could send me the links of videos it will be more helpful.
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Sorry Ana... I had forgotton to thank you. But I had watched the video already.
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I'm attempting to carry out some experiments which require me to remove long stretches of intact spinal cord.  I came across some literature which describes hydraulic extraction using a syringe to push the entire spinal cord out in one shot.  I have tried a few times but the cord doesn't seem to come out as easily/cleanly as the literature suggests.  Any advice on how to improve my extractions?
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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
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I am working on some tissues (Liver, Spleen, Pancreas) to isolate nuclei, every time I try to isolate the nuclei, the sample full of red blood cells and I can't distinguish between nuclei and RBC. Is there any way to eliminate the RBC?
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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.
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I found out it's about prevention of damaging the nerve tissue, etc. from google but I would like further precise explanation for this please. Thank you.
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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.
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Recently the University Grants Commission, New Delhi, issued a directive to all colleges and universities in the country to find alternatives to animal dissection and animal experimentation especially at undergraduate and post-graduate levels. I want to know if some is using such alternatives and if that could be shared?
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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.
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I need to perform simultaneous hippocampal and striatal dissections, is this possible without disrupting the tissue? The mice are 12 months old. Thank you very much!
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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
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I want to micro dissect the brain of rats and mice to isolate hippocampus, prefrontal cortex and other brain parts. Can help me out to find the best video on micro dissection of brain either in youtube or any other website? Thank you.
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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!
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I am trying to collect the hippocampus, prefrontal cortex and amygdala from adolescent rat brains (unfixed).  Is there a way I could validate the accuracy of the dissection to rule out contamination from adjacent brain regions? For instance, I have been searching for molecular markers of the hippocampus that are absent in adjacent regions, and vice versa.
Thank you
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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
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I am finding it difficult to identify the lymph node. I would like to remove then, paraffin embed and stain with H&E.
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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
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I am wondering if there is a method for dissecting adjacent normal and tumor samples from living mice without sacrificing the mouse? I hope to study the tumor evolution at multiple time points from the same mice. Is it possible?
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Thanks! It is what I am looking for!