A Minimally-Invasive Method for Serial Cerebrospinal Fluid Collection and Injection in Rodents with High Survival Rates

Abstract

Cerebrospinal fluid (CSF) is an important sample source for diagnosing diseases in the central nervous system (CNS), but collecting and injecting CSF in small animals is technically challenging and often results in high mortality rates. Here, we present a cost-effective and efficient method for accessing the CSF in live rodents for fluid collection and infusion purposes. The key element of this protocol is a metal needle tool bent at a unique angle and length, allowing the successful access of the CSF through the foramen magnum. With this method, we can collect 5–10 µL of the CSF from mice and 70–100 µL from rats for downstream analyses, including mass spectrometry. Moreover, our minimally-invasive procedure enables iterative CSF collection from the same animal every few days, representing a significant improvement over prior protocols. Additionally, our method can be used to inject solutions into mice cisterna magna with high success rates and high postoperative recovery rates. In summary, we provide an efficient and minimally-invasive protocol for collecting and infusing reagents into the CSF in live rodents. We envision this protocol will facilitate biomarker discovery and drug development for diseases in the central nervous system.

Minimally invasive serial collection of cerebrospinal fluid reveals sex-dependent differences in neuroinflammation in a rat model of mild traumatic brain injury

“…CSF proteomics are not widely studied in pre-clinical models due to the difficult nature of CSF collection. Previously described methods of CSF collection do not allow for repeated collections, require specialized tools and equipment, and can cause further brain injury or death 14,15,37 . The serial collection method used herein did not cause the rats to experience unusually significant weight changes, exhibit evidence of pain/distress (shallow or labored breathing, poor grooming, hunching, or porphyria), show signs of infection or swelling, or die.…”

“…This protocol is adapted from Han et al making the procedure easy to adapt for any laboratory without the use of custom built equipment or specialized tools 15 . To prep for CSF collection, an isofluorane nose cone was taped to the short side of a rectangular brick (can be any raised platform such as a pipette tip box) so the rat's head could be positioned vertically while breathing in isofluorane.…”

Exosomes Derived from Bone Marrow Mesenchymal Stem Cells Alleviate Rheumatoid Arthritis Symptoms via Shuttling Proteins

“…The protein sample was loaded onto a 12% SDS-PAGE gel electrophoresis system, followed by staining using Coomassie brilliant blue R-250 and decolorization until the bands were distinctly visible. Next, each sample underwent trypsin treatment according to the previous steps …”

“…UHPLC-MS/MS analysis was conducted by Novogene Co., Ltd. Company (Beijing, China) using a nanoElute UHPLC system (Bruker, Germany) coupled with the tims TOF Pro 2 mass spectrometer (Bruker, Germany) as previously reported. 31 In other words, each sample was injected into the LC-MS system to detect. The analytical column utilized in the study was a column measuring 15 cm in length and 100 μm in diameter with a particle size of 1.9 μm.…”

Microfluidic interfaces for chronic bidirectional access to the brain

“…The gold-standard for collecting cerebrospinal fluid (CSF) in preclinical research is cisterna magna puncturing, an invasive procedure that grants ventricular access only once every two/three months 6,7 and has a high risk for blood contamination, which yields non-representative samples. Chronic cannulation approaches are technically demanding, do not provide direct access to the ventricle and their long-term stability has not been well characterized [8][9][10][11] . Microdialysis 12,13 and cerebral open flow microperfusion (cFOM) 14 on the other hand, cause significant tissue damage 15 due to their relatively large size (~0.5mm in diameter) and provide highly diluted samples which necessitate advanced downstream analytical methods.…”