with reference to various research articles; relating to various spinal cord injury experiments, the spinal cord is preferred to be harvested from live animal (mouse, dog) for the purpose of western blotting and immuno cytochemistry, but circumstances are not every time favorable to collect the spinal cord from live animal. do the livability of animal affect the integrity of spinal cord? can it be harvested while the animal is dead during experimental period?
i need to know the significance of spinal cord harvesting from live animal.
Kelly Bisciotti thanks for the answer but sampling from dead animal could affect the protein contents interms of inflammatory and neural markers?
neural markers (gfap, tuj1, NFM, GALC) and inflammatory markers (COX2, pstat, il6 etc). recently i compared dead and fresh samples and i got similar results.
The main puropose to investigate complete and fresh neural systems should be to learn more of the flexibility and other neuromechanical and neurodynamical characteristics .
In the period 1960-1985 the neuroscientist and neuroradiologist Milan Roth investigated the neuro-osseous growth realtions, that can be disturbed by internal factors ( lack of O2 at birth, poison, drugs) but it offers a good explantion too, how the sedenatary lifestyle of children hinders this groth by stretch of the CNS and priovide the western world with bad postures , early and seroious degenration, but also ( from own observations) very thight central cords that look responsble for Arnold Chiari, syringomeylie and : skeletal deformations as scoliosis. A spinal fracture will give central cord injuries much easier as the cord is thight already.
Roth did fsh cadaveric studies to show that the cord stays in central position in bending the spine in a young healthy boy, killed by accident ( to show the imortance of Holzer's Neuroprotective Mechanism ) In modern youth this would show inevitably a cord that will be pulled against the wall, if it is not already attached to it ( you can see this on many MRI's )
Focus on biochemic substances blurs the more important neuromechanic characteristics that make cords vulnerable for overstretch. If injures, the both parts of the cord will retract immediately, so sponteanous repair is impossible.
I intend to do histochemical staining on mice spinal cord.I tried to isolate the cord by cutting through vertebral column.Though its possible,it seems to take a lot of time,and injures spinal cord a lot.
Is it right to do IHC on spinal cord taken by the process of ejection?
Nia lopez, No for IHC it is very important to harvest spinal cord by laminectomy bcz ejection inure and deteriorate the injured segment and is no longer suitable for IHC.\
plz refer to this article
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3770983/
Do laminectomy, completely expose the desired spinal cord segment and cut away with scalpel. after harvesting put the spinal cord in 10% sucrose solution for 24 hrs and then 20% sucrose for 48 hrs. then embedd or freeze in OCT compund. after embedding in oct compund your sample is ready to cryosection at -20C. hope it understand you well.
dear Nia Lopez, can you plz elaborate your Question, it seem difficult for me to understand.
as i know if your r extracting mRNA in a sufficient quantity, using 1000ng in cDNA making and then running PCR.
set threshold values to 0.8 for all the number of variable/genes before saving in to the computer.
use GAPDH as house keeping.
i hope it will work, or better to revise your experiment and identify where you r doing the mistake.
good luck
- Badges
- Science topic
- Similar topics
- Analytical Chemistry
- Extraction