Name: Rapid Isolation of Dorsal Root Ganglion Macrophages (Video) | JoVE Journal
Uploaded: 2019-09-07T14:30:00.000+00:00
Duration: 7 min 22 s
Description: Learn a rapid mechanical dissociation protocol for isolating dorsal root ganglion macrophages from mice. This enzyme-free method enables efficient phenotyping and fluorescence-activated cell sorting analysis for neuroimmune studies.

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5 minutes

The study was supported by: Foundation for Anesthesia Education and Research (XY); the UCSF Department of Anesthesia and Perioperative Care (XY); and 1R01NS100801-01(GZ). This study was supported in part by HDFCCC Laboratory for Cell Analysis Shared Resources Facility through a grant from the NIH (P30CA082103).

All animal experiments were approved by the Institutional Animal Care and Use Committee at the University of California San Francisco and were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals.
1. Collect lumbar DRG from experimental mice
<ol>
	<li>Before starting the experiment, prepare the working solution of the density gradient medium (e.g., Percoll) by mixing 9 volumes of the medium with 1 volume of Ca++/Mg++-free 10x HBSS. Keep it on ice.</li>
	<li>Anesthetize the mouse with 2.5% Avertin. Confirm that the animal is fully anesthetized by the lack of response to the hind paw pinch. 
	NOTE: Both male and female mice aged 6-8 weeks were used.</li>
	<li>Perfuse the mouse transcardially with 10 mL of pre-chilled 1x PBS.
	<ol>
		<li>Place the mouse in the supine position with four paws secured with the tape inside a chemical fume hood. Lift the skin below the rib cage by the forceps, and make a small incision with surgical scissors to expose the liver and diaphragm.</li>
		<li>Continue to use scissors to cut the diaphragm and the rib cage, open the pleural cavity to expose the beating heart.</li>
		<li>Quickly cut the right atrial appendage with iris scissors. Once the bleeding is noted, insert a 30 G needle into the posterior end of the left ventricle, and slowly inject 10 mL of pre-chilled 1x PBS to perfuse the animal.</li>
	</ol>
	</li>
	<li>Perform dorsal laminectomy15 on the mouse placed at the prone position. 
	NOTE: If cell culture is planned, spray the mouse with 70% ethanol before incision and use pre-sterilized surgical instruments for dissection.
	<ol>
		<li>Use a size 11 scalpel to make one longitudinal lateral deep incisions starting from the thoracic region down to the sacral region. Remove the skin by the scissors to expose the dorsal muscle layer.</li>
		<li>Use Friedman-Pearson Rongeur to peel off the connective tissues and muscles until the lumbosacral spine processes and bilateral transverse processes are exposed.</li>
		<li>Use a Noyes Spring Scissor to carefully open the dorsal spinal column, then switch to a Friedman-Pearson Rongeur to remove the vertebral bones to expose the spinal cord with intact spinal nerves attached.</li>
	</ol>
	</li>
	<li>Carefully dissect ipsilateral and contralateral lumbar DRG (L4/L5 DRG in our study) and place it into 1 mL of ice-cold Ca++/Mg++-free 1x HBSS in a Dounce tissue homogenizer. Now the tissues are ready for step 2. 
	NOTE: Trim the spinal nerve attached to the DRG if possible.</li>
</ol>
2. Isolate single cells from mouse lumbar DRG
<ol>
	<li>Homogenize the DRG tissue with a loose pestle in the Dounce homogenizer&#160;20-25 times.</li>
	<li>Place a sterile 70 &#956;m nylon cell strainer in a sterile 50 mL conical tube. Wet the cell strainer with 800 &#956;L of ice-cold 1x HBSS, and the flow-through is collected in the conical tube.</li>
	<li>Collect the homogenized tissue suspension from the homogenizer using a pipette and pass through the wet 70 &#956;m nylon cell strainer into the 50 mL conical tube.</li>
	<li>Rinse the homogenizer twice with 800 &#956;L of ice-cold 1x HBSS and then decant into the same 50 mL conical tube with cell strainer to increase the yield.</li>
	<li>Add 1.5 mL of equilibrated ice-cold isotonic density gradient medium (prepared in step 1.1) into a sterile 5-mL polystyrene FACS tube.</li>
	<li>Transfer the cell homogenate from the 50 mL conical tube (in step 2.4) into the FACS tube, mix well with the density gradient medium by pipetting up and down. Add an additional 500 &#956;L of 1x HBSS to seal the top.</li>
	<li>Pellet the cells by centrifugation at 800 x g for 20 min at 4 &#176;C.</li>
	<li>Carefully aspirate the supernatant containing myelin in the medium without disturbing the cell pellet at the bottom of the FACS tube.</li>
	<li>Resuspend the cells in PBS or FACS buffer containing 5% Fetal bovine serum (FBS) for FACS analysis. 
	NOTE: At least 50,000 to 100,000 cells are expected from L4 /L5 DRG of one mouse.
	<ol>
		<li>Resuspend the mechanically isolated DRG cells (L4/L5) in 100 &#956;L of PBS containing 5% Fetal Bovine Serum and then incubate with &#945;-mouse CX3CR1-APC antibody (1:2,000) in the dark at 4 &#176;C for 1 h.</li>
		<li>Wash the cells with 5 mL of PBS once; centrifuge the cells 360 x g for 8 min at 4 &#176;C.</li>
		<li>Aspirate the supernatant, then resuspend the cell pellet in 300 &#956;L of PBS for FCAS analysis. If cell sorting is planned, resuspend the cells in the FACS buffer instead.</li>
	</ol>
	</li>
</ol>

<ol><li>Ji, R. R., Chamessian, A., Zhang, Y. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pain+regulation+by+non-neuronal+cells+and+inflammation%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Pain regulation by non-neuronal cells and inflammation.</a> Science. 354 (6312), 572-577 (2016).</li>
<li>Inoue, K., Tsuda, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microglia+in+neuropathic+pain%3A+cellular+and+molecular+mechanisms+and+therapeutic+potential%5BTitle%5D)+AND+%22Nature+Reviews+Neurosciences%22%5BJournal%5D)">Microglia in neuropathic pain: cellular and molecular mechanisms and therapeutic potential.</a> Nature Reviews Neurosciences. 19 (3), 138-152 (2018).</li>
<li>Hu, P., McLachlan, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Distinct+functional+types+of+macrophage+in+dorsal+root+ganglia+and+spinal+nerves+proximal+to+sciatic+and+spinal+nerve+transections+in+the+rat%5BTitle%5D)+AND+%22Experimental+Neurology%22%5BJournal%5D)">Distinct functional types of macrophage in dorsal root ganglia and spinal nerves proximal to sciatic and spinal nerve transections in the rat.</a> Experimental Neurology. 184 (2), 590-605 (2003).</li>
<li>Simeoli, R., Montague, K., Jones, H. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Exosomal+cargo+including+microRNA+regulates+sensory+neuron+to+macrophage+communication+after+nerve+trauma%5BTitle%5D)+AND+%22Nature+Communication%22%5BJournal%5D)">Exosomal cargo including microRNA regulates sensory neuron to macrophage communication after nerve trauma.</a> Nature Communication. 8 (1), 1778(2017).</li>
<li>Cobos, E. J., Nickerson, C. A., Gao, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mechanistic+differences+in+neuropathic+pain+modalities+revealed+by+correlating+behavior+with+global+expression+profiling%5BTitle%5D)+AND+%22Cell+Reports%22%5BJournal%5D)">Mechanistic differences in neuropathic pain modalities revealed by correlating behavior with global expression profiling.</a> Cell Reports. 22 (5), 1301-1312 (2018).</li>
<li>Zhang, H., Li, Y., de Carvalho-Barbosa, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dorsal+Root+Ganglion+Infiltration+by+Macrophages+Contributes+to+Paclitaxel+Chemotherapy-Induced+Peripheral+Neuropathy%5BTitle%5D)+AND+%22The+Journal+of+Pain%22%5BJournal%5D)">Dorsal Root Ganglion Infiltration by Macrophages Contributes to Paclitaxel Chemotherapy-Induced Peripheral Neuropathy.</a> The Journal of Pain. 17 (7), 775-786 (2016).</li>
<li>Liu, T., van Rooijen, N., Tracey, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Depletion+of+macrophages+reduces+axonal+degeneration+and+hyperalgesia+following+nerve+injury%5BTitle%5D)+AND+%22Pain%22%5BJournal%5D)">Depletion of macrophages reduces axonal degeneration and hyperalgesia following nerve injury.</a> Pain. 86 (1-2), 25-32 (2000).</li>
<li>Peng, J., Gu, N., Zhou, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microglia+and+monocytes+synergistically+promote+the+transition+from+acute+to+chronic+pain+after+nerve+injury%5BTitle%5D)+AND+%22Nature+Communication%22%5BJournal%5D)">Microglia and monocytes synergistically promote the transition from acute to chronic pain after nerve injury.</a> Nature Communication. 7, 12029(2016).</li>
<li>Barclay, J., Clark, A. K., Ganju, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Role+of+the+cysteine+protease+cathepsin+S+in+neuropathic+hyperalgesia%5BTitle%5D)+AND+%22Pain%22%5BJournal%5D)">Role of the cysteine protease cathepsin S in neuropathic hyperalgesia.</a> Pain. 130 (3), 225-234 (2007).</li>
<li>Lim, H., Lee, H., Noh, K., Lee, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((IKK%2FNF-kappaB-dependent+satellite+glia+activation+induces+spinal+cord+microglia+activation+and+neuropathic+pain+after+nerve+injury%5BTitle%5D)+AND+%22Pain%22%5BJournal%5D)">IKK/NF-kappaB-dependent satellite glia activation induces spinal cord microglia activation and neuropathic pain after nerve injury.</a> Pain. 158 (9), 1666-1677 (2017).</li>
<li>Rutkowski, M. D., Pahl, J. L., Sweitzer, S., van Rooijen, N., DeLeo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Limited+role+of+macrophages+in+generation+of+nerve+injury-induced+mechanical+allodynia%5BTitle%5D)+AND+%22Physiology+and+Behavior%22%5BJournal%5D)">Limited role of macrophages in generation of nerve injury-induced mechanical allodynia.</a> Physiology and Behavior. (3-4), 225-235 (2000).</li>
<li>Kwon, M. J., Shin, H. Y., Cui, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CCL2+Mediates+Neuron-Macrophage+Interactions+to+Drive+Proregenerative+Macrophage+Activation+Following+Preconditioning+Injury%5BTitle%5D)+AND+%22Journal+of+Neuroscience%22%5BJournal%5D)">CCL2 Mediates Neuron-Macrophage Interactions to Drive Proregenerative Macrophage Activation Following Preconditioning Injury.</a> Journal of Neuroscience. 35 (48), 15934-15947 (2015).</li>
<li>Zigmond, R. E., Echevarria, F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Macrophage+biology+in+the+peripheral+nervous+system+after+injury%5BTitle%5D)+AND+%22Progress+in+Neurobiology%22%5BJournal%5D)">Macrophage biology in the peripheral nervous system after injury.</a> Progress in Neurobiology. 173, 102-121 (2019).</li>
<li>Ghasemlou, N., Chiu, I. M., Julien, J. P., Woolf, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CD11b%2BLy6G-+myeloid+cells+mediate+mechanical+inflammatory+pain+hypersensitivity%5BTitle%5D)+AND+%22Proceedings+of+the+National+Academy+of+Science+USA%22%5BJournal%5D)">CD11b+Ly6G- myeloid cells mediate mechanical inflammatory pain hypersensitivity.</a> Proceedings of the National Academy of Science USA. 112 (49), E6808-E6817 (2015).</li>
<li>Malin, S. A., Davis, B. M., Molliver, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Production+of+dissociated+sensory+neuron+cultures+and+considerations+for+their+use+in+studying+neuronal+function+and+plasticity%5BTitle%5D)+AND+%22Nature+Protocols%22%5BJournal%5D)">Production of dissociated sensory neuron cultures and considerations for their use in studying neuronal function and plasticity.</a> Nature Protocols. 2 (1), 152-160 (2007).</li>
<li>Lin, Y. T., Chen, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dorsal+Root+Ganglia+Isolation+and+Primary+Culture+to+Study+Neurotransmitter+Release%5BTitle%5D)+AND+%22Journal+of+Visualized+Experiment%22%5BJournal%5D)">Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release.</a> Journal of Visualized Experiment. (140), (2018).</li>
<li>Burnett, S. H., Kershen, E. J., Zhang, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Conditional+macrophage+ablation+in+transgenic+mice+expressing+a+Fas-based+suicide+gene%5BTitle%5D)+AND+%22J+Leukoc+Biol%22%5BJournal%5D)">Conditional macrophage ablation in transgenic mice expressing a Fas-based suicide gene.</a> J Leukoc Biol. 75 (4), 612-623 (2004).</li>
<li>Lopes, D. M., Malek, N., Edye, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sex+differences+in+peripheral+not+central+immune+responses+to+pain-inducing+injury%5BTitle%5D)+AND+%22Science+Reports%22%5BJournal%5D)">Sex differences in peripheral not central immune responses to pain-inducing injury.</a> Science Reports. 7 (1), 16460(2017).</li>
<li>Kim, Y. S., Anderson, M., Park, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Coupled+Activation+of+Primary+Sensory+Neurons+Contributes+to+Chronic+Pain%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">Coupled Activation of Primary Sensory Neurons Contributes to Chronic Pain.</a> Neuron. 91 (5), 1085-1096 (2016).</li>
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The authors declare no competing financial interests related to this manuscript.

Here we introduce a new method to effectively enrich isolated macrophages from mouse DRG. The conventional approach to isolate DRG immune cells requires enzymatic digestion15,18, which is now replaced with mechanical homogenization in our protocol to limit undesired cell damage and increase the yield. Therefore, the new protocol is far less time consuming. More importantly, enzyme digestion might stimulate the macrophages and change the molecular signature. In co...

There is now considerable evidence that immune cells contribute to the neuropathic pain following peripheral nerve injury1,2. Peripheral monocytic cells, including mature macrophages, are known to respond to tissue injury and systemic infection through phagocytosis, antigen presentation, and cytokine release. Paralleling the nerve injury-induced microglia activation in the spinal dorsal horn, macrophages in the dorsal root ganglia (DRG) also expand significantly after nerve injury3,4. Notably, there are growing interests to determine if macrophages contribute to neuropathic pain development after peripheral nerve injury by interacting with sensory neurons in the DRG5,6,7,8,9,10,11. Moreover, recent studies also implicate the contribution of DRG macrophages in the axonal repair after nerve injury12,13. Another study further suggests that macrophage subpopulations (i.e., CD11b+Ly6Chi and CD11b+Ly6Clow/- cells) may play a different role in the mechanical hypersensitivity14. Therefore, rapidly phenotyping the response of DRG macrophages in the context of nerve injury may help us identify neuroimmune factors contributing to neuropathic pain.
Conventionally, the protocol to isolate macrophages in the DRG involves multiple steps including enzymatic digestion15,16. The technique is often time consuming and can be costly for large-scale experiments. Although mild digestion with collagenase type II (4 mg/mL) and dispase type II (4.7 mg/mL) for 20 min was recommended previously15, it is conceivable that cells after the exposure to this enzyme are prone to cell damage or cell death, which may lead to low yield. In addition, the difference in the quality of enzymes from batch to batch may further impact the efficiency of this process. More importantly, macrophages exposed to the enzyme digestion might be undesirably stimulated and thus can be very different from the in-vivo status. The changes may potentially complicate the outcome of the functional study.
Here we describe an enzyme-free protocol to rapidly isolate DRG macrophages at 4 &#176;C using mechanical dissociation. The samples are kept on ice to limit cellular stress. As a result, our approach provides an advantage to maintain consistency of the isolation, and the isolated cells are presumably healthier and less stimulated. We further present the evidence to validate the quality of the isolated cells with Fluorescence-activated Cell Sorting (FACS) analysis.

<table><tbody><tr><td>AP20187</td><td>Clontech</td><td>635058</td><td></td></tr><tr><td>&amp;#945;-mouse CX3CR1-APC antibody</td><td>Biolegend</td><td>149007</td><td></td></tr><tr><td>Avertin</td><td>Sigma</td><td>T48402</td><td></td></tr><tr><td>Cell strainer (70 mm nylon)</td><td>Falcon</td><td>352350</td><td></td></tr><tr><td>Centrifuge</td><td>Eppendorf</td><td>5810R</td><td></td></tr><tr><td>Dounce tissue homogenizer</td><td>Wheaton</td><td>357538 (1ml)</td><td></td></tr><tr><td>FACS tubes (5ml)</td><td>Falcon</td><td>352052</td><td></td></tr><tr><td>Friedman-Pearson Rongeur</td><td>FST</td><td>16121-14</td><td></td></tr><tr><td>HBSS (10x, Ca++/Mg++-free)</td><td>Gibco</td><td>14185-052</td><td></td></tr><tr><td>Noyes Spring Scissor</td><td>FST</td><td>15012-12</td><td></td></tr><tr><td>Percoll</td><td>Sigma</td><td>P4937-500ml</td><td></td></tr><tr><td>Propidium iodide</td><td>Sigma</td><td>P4864-10ml</td><td></td></tr></tbody></table>

rapid isolation of dorsal root ganglion macrophages

Rapid Isolation of Dorsal Root Ganglion Macrophages

There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after peripheral nerve injury. Peripheral monocytic cells, including macrophages, are known to respond to a tissue injury through phagocytosis, antigen presentation, and cytokine release. Emerging evidence has implicated the contribution of dorsal root ganglia macrophages to neuropathic pain development and axonal repair in the context of nerve injury. Rapidly phenotyping (or &#8220;rapid isolation of&#8221;) the response of dorsal root ganglia macrophages in the context of nerve injury is desired to identify the unknown neuroimmune factors. Here we demonstrate how our lab rapidly and effectively isolates macrophages from the dorsal root ganglia using an enzyme-free mechanical dissociation protocol. The samples are kept on ice throughout to limit cellular stress. This protocol is far less time consuming compared to the standard enzymatic protocol and has been routinely used for our Fluorescence-activated Cell Sorting analysis.

To validate the isolated cells, we first chose the Macrophage Fas-Induced Apoptosis (MAFIA) transgenic mice17. This line expresses a drug-inducible FK506-binding protein (FKBP)-Fas suicide fusion gene and green fluorescent protein (eGFP) under the control of the promoter of CSF1 receptor (CSF1R), which is specifically expressed in both macrophages and microglia. Systemic injection of FK-binding protein dimerizer, AP20187 (AP), induces the apoptosis of the cells expressing the transgene. The expres...

Here we present a mechanical dissociation protocol to rapidly isolate macrophages from the dorsal root ganglion for phenotyping and functional analysis.

There are growing interests to study the molecular and cellular interactions among immune cells and sensory neurons in the dorsal root ganglia after ...

rapid-isolation-of-dorsal-root-ganglion-macrophages

Research

JoVE Journal

Neuroscience

This video demonstrates an enzyme-free protocol to isolate macrophages from mouse dorsal root ganglion (DRG) using a mechanical dissociation technique. Upon homogenization of the freshly collected DRG tissue, antibodies against the cell-surface receptors on the macrophages are used to label the cells fluorescently, followed by performing flow cytometry to confirm the presence of macrophages in the sample.

Isolation of Macrophages from Mouse Dorsal Root Ganglion Using Mechanical Dissociation

This video demonstrates a method for harvesting neurons from the dorsal root ganglia (DRG). DRG from a rat's spinal column is isolated, treated with enzymes, and then mechanically dissociated to obtain a single-cell suspension. The neurons are separated from glial cells using density gradient centrifugation and cultured in a growth medium.

A Technique for Harvesting and Dissociating Dorsal Root Ganglia for Neuronal Cultures

This video demonstrates a protocol for enriching neuronal cultures from dorsal root ganglia by immunopanning, thereby improving their purity and utility for neurological research.

Neuronal Enrichment of Dorsal Root Ganglia by Immunopanning

This video demonstrates a technique for isolating and culturing dorsal root ganglia (DRG) cells from the rat spinal column. DRGs excised from the spinal column are treated with enzymes and mechanically dissociated to obtain a single-cell suspension. The cells are cultured in a medium containing inhibitors and growth factors that limit glial cell proliferation and promote neuronal growth, respectively.

Establishing a Primary Dorsal Root Ganglia Cell Culture from a Rat Spinal Column

Dorsal root ganglia (DRG) are structures containing the sensory neurons of the peripheral nervous system. When dissociated, they can be co-cultured with SC-like adipose-derived stem cells (ASC), providing a valuable model to study in vitro nerve regeneration and myelination, mimicking the in vivo environment at the injury site.

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

This protocol describes the isolation of dorsal root ganglion (DRG) neurons isolated from rats and the culture of DRG neurons on a static pre-stretched cell culture system to enhance axon alignment, with subsequent co-culture of Schwann Cells (SCs) to promote myelination.

An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons

Here, we present a protocol for hydraulic extrusion of the spinal cord as well as identification and isolation of specific dorsal root ganglia (DRGs) in the same rodent. Compared to standard spinal cord isolation methods, this method is significantly faster and reduces the risk of tissue damage.

Hydraulic Extrusion of the Spinal Cord and Isolation of Dorsal Root Ganglia in Rodents

Efforts to understand microglial function in detail have been hindered by the lack of microglial culture models that recapitulate the properties of mature in vivo microglia. This protocol describes an isolation and culture approach designed to maintain robust survival of highly ramified mature rat microglia under defined-medium conditions.

Isolation and Culture of Rodent Microglia to Promote a Dynamic Ramified Morphology in Serum-free Medium

Dorsal root ganglia (DRG) primary cultures are frequently used to study physiological functions or pathology-related events in sensory neurons. Here, we demonstrate the use of lumbar DRG cultures to detect the release of neurotransmitters after neuropeptide FF receptor type 2 stimulation with a selective agonist.

Dorsal Root Ganglia Isolation and Primary Culture to Study Neurotransmitter Release

This protocol provides a technique to harvest and culture explanted dorsal root ganglion (DRG) from adult Sprague Dawley rats in a multi-compartment (MC) device.

Harvesting, Embedding, and Culturing Dorsal Root Ganglia in Multi-compartment Devices to Study Peripheral Neuronal Features

Dorsal root ganglia (DRG) contain primary sensory neuron cell bodies and relay peripheral sensory signals to the spinal cord. This protocol describes a method for mouse DRG dissection that supports standardized isolation of structurally intact ganglia, anatomical segmental identification, intuitive guidance for novice users, and reduced procedural tissue injury.

Dissection and Isolation of Mouse Dorsal Root Ganglia

Learn a rapid mechanical dissociation protocol for isolating dorsal root ganglion macrophages from mice. This enzyme-free method enables efficient phenotyping and fluorescence-activated cell sorting analysis for neuroimmune studies.

Rapid Isolation of Dorsal Root Ganglion Macrophages

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