# Experimental Techniques **URL:** https://forum.painresearcher.net/c/experimental-techniques/6.md [Latest](https://forum.painresearcher.net/latest.md) · [Categories](https://forum.painresearcher.net/categories.md) · [Tags](https://forum.painresearcher.net/tags.md) --- ## [About the Experimental Techniques category](https://forum.painresearcher.net/t/about-the-experimental-techniques-category/15)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 1\ **Last updated:** [December 12, 2017, 1:05am UTC](https://forum.painresearcher.net/t/about-the-experimental-techniques-category/15 "2017-12-12T01:05:24Z")
This category is for discussing the technical details of pain experiments. --- ## [Probelm with Patch clamp of Dissociated DRG neurons](https://forum.painresearcher.net/t/probelm-with-patch-clamp-of-dissociated-drg-neurons/822)
**Author:** [@Abderaouf](https://forum.painresearcher.net/u/Abderaouf)\ **Replies:** 7\ **Last updated:** [September 17, 2026, 6:50pm UTC](https://forum.painresearcher.net/t/probelm-with-patch-clamp-of-dissociated-drg-neurons/822 "2026-09-17T18:50:14Z")
Hi everyone We are trying to study the electrophysiological parameters in dissociated DRG primary culture cells, we have been trying for a while now to get a whole cell configuration to do a patch clamp experiments but … --- ## [Alternative to TUBB3 (TUJ1) for axon staining, non-neuronal staining](https://forum.painresearcher.net/t/alternative-to-tubb3-tuj1-for-axon-staining-non-neuronal-staining/844)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 2\ **Last updated:** [April 13, 2026, 3:27pm UTC](https://forum.painresearcher.net/t/alternative-to-tubb3-tuj1-for-axon-staining-non-neuronal-staining/844 "2026-04-13T15:27:25Z")
Hi all, I’m looking at axons from mouse and human DRG in vitro. The standard stain is TUBB3. I’m using the Tuj-1 clone from Biolegend (formerly Covaris), which is the gold-standard as far as I can tell. https://www.bio… --- ## [Immunostaining of paraffin-embedded tissues with bone](https://forum.painresearcher.net/t/immunostaining-of-paraffin-embedded-tissues-with-bone/1453)
**Author:** [@wtheodoro](https://forum.painresearcher.net/u/wtheodoro)\ **Replies:** 2\ **Last updated:** [November 4, 2025, 2:05pm UTC](https://forum.painresearcher.net/t/immunostaining-of-paraffin-embedded-tissues-with-bone/1453 "2025-11-04T14:05:39Z")
Hello everyone, I’ve been encountering high background when immunostaining paraffin-embedded, fixed tissues that include both soft and bony components. These specimens are periapical tissues obtained from human teeth d… --- ## [RNAscope (In situ hybridization) to study gene expression in pain-related tissues](https://forum.painresearcher.net/t/rnascope-in-situ-hybridization-to-study-gene-expression-in-pain-related-tissues/102)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 88\ **Last updated:** [October 23, 2025, 2:43pm UTC](https://forum.painresearcher.net/t/rnascope-in-situ-hybridization-to-study-gene-expression-in-pain-related-tissues/102 "2025-10-23T14:43:54Z")
In situ hybridization is a powerful method to interrogate gene expression in tissues. It is known as being a challenging procedure due to the many steps where errors and obstacles arise. We've recently begun to use a com… --- ## [Pain behaviors in face](https://forum.painresearcher.net/t/pain-behaviors-in-face/1451)
**Author:** [@Malaika](https://forum.painresearcher.net/u/Malaika)\ **Replies:** 0\ **Last updated:** [October 15, 2025, 6:44pm UTC](https://forum.painresearcher.net/t/pain-behaviors-in-face/1451 "2025-10-15T18:44:57Z")
Does anyone have any recommendations on what good programs there are to track facial pain behaviors specifically things like whisking, face wiping, grimacing, etc? I know Pain Face is one that many use but it seems to no… --- ## [Tone-pain conditioning](https://forum.painresearcher.net/t/tone-pain-conditioning/1443)
**Author:** [@Malaika](https://forum.painresearcher.net/u/Malaika)\ **Replies:** 2\ **Last updated:** [September 25, 2025, 6:09pm UTC](https://forum.painresearcher.net/t/tone-pain-conditioning/1443 "2025-09-25T18:09:59Z")
Has anyone done a tone-pain conditioning task specifically using a laser to administer pain in mice? I’m curious to know the parameters (i.e. how long tone was played for, what intensity of the laser was used, what kind … --- ## [Human on Human high IgG background on human DRG/nerve](https://forum.painresearcher.net/t/human-on-human-high-igg-background-on-human-drg-nerve/784)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 4\ **Last updated:** [September 5, 2025, 12:19pm UTC](https://forum.painresearcher.net/t/human-on-human-high-igg-background-on-human-drg-nerve/784 "2025-09-05T12:19:00Z")
I’m attempting IHC on human DRG/nerve using human serum (containing putative anti-human antibodies). The issue I’m running into is that there is huge background with just the secondary alone (goat anti-human IgG). How … --- ## [EF1a promoter to drive expression in DRG in vivo?](https://forum.painresearcher.net/t/ef1a-promoter-to-drive-expression-in-drg-in-vivo/331)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 14\ **Last updated:** [August 19, 2025, 3:56pm UTC](https://forum.painresearcher.net/t/ef1a-promoter-to-drive-expression-in-drg-in-vivo/331 "2025-08-19T15:56:49Z")
Has anyone used EF1a promoters to drive transgene expression in DRGs in vivo? I’ve had success with hSyn, and recently, a paper from Haenraets et al showed CAG works well for DRG. But EF1a is also very popular and it’d b… --- ## [Inquiry on NR2B Primary Antibody for Mouse Fluorescent Immunostaining](https://forum.painresearcher.net/t/inquiry-on-nr2b-primary-antibody-for-mouse-fluorescent-immunostaining/1432)
**Author:** [@ZoeChen](https://forum.painresearcher.net/u/ZoeChen)\ **Replies:** 1\ **Last updated:** [August 14, 2025, 3:07pm UTC](https://forum.painresearcher.net/t/inquiry-on-nr2b-primary-antibody-for-mouse-fluorescent-immunostaining/1432 "2025-08-14T15:07:25Z")
Dear all, I am a graduate student currently working on fluorescent immunostaining of mice spinal cord tissue. I am searching for a primary antibody against NR2B that is suitable for immunofluorescence and raised in a ho… --- ## [Transgenic Cre mouse for labeling periphery nerves](https://forum.painresearcher.net/t/transgenic-cre-mouse-for-labeling-periphery-nerves/1398)
**Author:** [@Jesicawiiiij](https://forum.painresearcher.net/u/Jesicawiiiij)\ **Replies:** 2\ **Last updated:** [August 11, 2025, 2:59pm UTC](https://forum.painresearcher.net/t/transgenic-cre-mouse-for-labeling-periphery-nerves/1398 "2025-08-11T14:59:07Z")
Hi everyone! I’m looking for Cre mouse line recommendations\*\* for comprehensive peripheral nerve labeling. My goal is to cross Cre mice with fluorescent reporters, then perform tissue clearing and whole-body peripheral n… --- ## [Request to provide a standard method of Mouse DRG culture and Immunocytochemistry Protocol](https://forum.painresearcher.net/t/request-to-provide-a-standard-method-of-mouse-drg-culture-and-immunocytochemistry-protocol/1415)
**Author:** [@Hemanta](https://forum.painresearcher.net/u/Hemanta)\ **Replies:** 0\ **Last updated:** [July 27, 2025, 6:37am UTC](https://forum.painresearcher.net/t/request-to-provide-a-standard-method-of-mouse-drg-culture-and-immunocytochemistry-protocol/1415 "2025-07-27T06:37:28Z")
I am searching for DRG culture and ICC protocol. --- ## [Housekeeping proteins for western blot that do not change following nerve injury](https://forum.painresearcher.net/t/housekeeping-proteins-for-western-blot-that-do-not-change-following-nerve-injury/1352)
**Author:** [@gg9761](https://forum.painresearcher.net/u/gg9761)\ **Replies:** 3\ **Last updated:** [July 9, 2025, 1:53pm UTC](https://forum.painresearcher.net/t/housekeeping-proteins-for-western-blot-that-do-not-change-following-nerve-injury/1352 "2025-07-09T13:53:29Z")
Hi all, I need to run a western blot on protein lysate extracted from mouse DRGs. One of my groups has undergone a spared nerve injury and I’m aware that certain commonly used housekeeping proteins can be up or downregu… --- ## [Ideal age to record PSCs from mice spinal cord without stimulation electrode](https://forum.painresearcher.net/t/ideal-age-to-record-pscs-from-mice-spinal-cord-without-stimulation-electrode/1359)
**Author:** [@anutoshroy](https://forum.painresearcher.net/u/anutoshroy)\ **Replies:** 2\ **Last updated:** [April 16, 2025, 4:10pm UTC](https://forum.painresearcher.net/t/ideal-age-to-record-pscs-from-mice-spinal-cord-without-stimulation-electrode/1359 "2025-04-16T16:10:32Z")
Hello! I have two queries regarding ephys from spinal cord slice, For recording of Post synaptic currents, is there any method other than stimulation electrode to record the PSCs in mice spinal cord in P0-P12 mice spi… --- ## [DAB staining Immunohistochemistery](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353)
**Author:** [@HebaReda](https://forum.painresearcher.net/u/HebaReda)\ **Replies:** 2\ **Last updated:** [April 6, 2025, 9:20pm UTC](https://forum.painresearcher.net/t/dab-staining-immunohistochemistery/1353 "2025-04-06T21:20:39Z")
Hi, I am planning to use DAB with nickel in immunohistochemistery so what is the best concentration that should be used please as I am confused with that? I was preparing it freshly by adding 2.7 mg in 10 ml PBS (so a … --- ## [Frozen vs. Non-frozen IHC (Fixed) Spinal Cords. Thoughts?](https://forum.painresearcher.net/t/frozen-vs-non-frozen-ihc-fixed-spinal-cords-thoughts/202)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 13\ **Last updated:** [March 17, 2025, 8:26pm UTC](https://forum.painresearcher.net/t/frozen-vs-non-frozen-ihc-fixed-spinal-cords-thoughts/202 "2025-03-17T20:26:46Z")
Has anyone done free-floating spinal cord sectioning/staining using fixed, but not frozen tissue using a vibratome? For example, I noticed Andrew Todd does much of his IHC this way, and the images look great. Any thought… --- ## [Establishment of a Magnetically Controlled Scalable Nerve Injury Model](https://forum.painresearcher.net/t/establishment-of-a-magnetically-controlled-scalable-nerve-injury-model/1282)
**Author:** [@PRF-Editorial-Board](https://forum.painresearcher.net/u/PRF-Editorial-Board)\ **Replies:** 0\ **Last updated:** [September 20, 2024, 7:18pm UTC](https://forum.painresearcher.net/t/establishment-of-a-magnetically-controlled-scalable-nerve-injury-model/1282 "2024-09-20T19:18:35Z")
What do you think of this new experimental method that provides quantitative and off-body controllable injury properties via a magnetically controlled clamp in rodent models? --- ## [Protein Isolation for Western Blots (protocols)?](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 3\ **Last updated:** [July 30, 2024, 6:12pm UTC](https://forum.painresearcher.net/t/protein-isolation-for-western-blots-protocols/785 "2024-07-30T18:12:41Z")
Hey all, Looking to do some protein work soon. I have some personal protocols from grad school but it’d be nice to see what people are doing. If you’re willing, share your protocol for: Mouse Spinal cord DRG Nerve H… --- ## [Pgp9.5 glabrous skin staining](https://forum.painresearcher.net/t/pgp9-5-glabrous-skin-staining/177)
**Author:** [@vanja](https://forum.painresearcher.net/u/vanja)\ **Replies:** 20\ **Last updated:** [July 10, 2024, 5:02am UTC](https://forum.painresearcher.net/t/pgp9-5-glabrous-skin-staining/177 "2024-07-10T05:02:33Z")
Hi all, could anyone recommend a good protocol for pgp9.5 staining of mouse foot pad skin? I get some dermal pgp9.5 staining, but never the small nerve endings that should enervate the epidermis. I have seen many differe… --- ## [P2X3 Antibodies](https://forum.painresearcher.net/t/p2x3-antibodies/1181)
**Author:** [@dmolliver](https://forum.painresearcher.net/u/dmolliver)\ **Replies:** 7\ **Last updated:** [June 19, 2024, 1:11pm UTC](https://forum.painresearcher.net/t/p2x3-antibodies/1181 "2024-06-19T13:11:10Z")
Can anyone recommend a good clean western blotting antibody for human P2X3? It looks like most of the widely-used antibodies for rodent P2X3 don’t work in human. --- ## [Are there dyes to label neurons, similar to how SR101 labels astrocytes in 20 minutes under 2P?](https://forum.painresearcher.net/t/are-there-dyes-to-label-neurons-similar-to-how-sr101-labels-astrocytes-in-20-minutes-under-2p/1201)
**Author:** [@Mengya](https://forum.painresearcher.net/u/Mengya)\ **Replies:** 0\ **Last updated:** [May 30, 2024, 9:30pm UTC](https://forum.painresearcher.net/t/are-there-dyes-to-label-neurons-similar-to-how-sr101-labels-astrocytes-in-20-minutes-under-2p/1201 "2024-05-30T21:30:09Z")
Hi, I am performing 2P imaging on the spinal cord using vGlut2-GCaMP mice. My goal is to calculate the percentage of responsive neurons to mechanical stimuli relative to the total number of neurons in the FOV. However, … --- ## [Intraganglionic injections into mouse DRG?](https://forum.painresearcher.net/t/intraganglionic-injections-into-mouse-drg/56)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 20\ **Last updated:** [May 8, 2024, 3:49pm UTC](https://forum.painresearcher.net/t/intraganglionic-injections-into-mouse-drg/56 "2024-05-08T15:49:15Z")
I'm needing to inject viral vectors directly into mouse DRG so that I can localize the expression just to the DRG. I can get very good expression when I inject intrathecally, but that enables the spread of the virus to p… --- ## [Optimizing Electronic Von Frey for Mouse](https://forum.painresearcher.net/t/optimizing-electronic-von-frey-for-mouse/1189)
**Author:** [@Anthony](https://forum.painresearcher.net/u/Anthony)\ **Replies:** 1\ **Last updated:** [April 11, 2024, 9:42pm UTC](https://forum.painresearcher.net/t/optimizing-electronic-von-frey-for-mouse/1189 "2024-04-11T21:42:47Z")
My project involves characterizing pain responses in mice following chronic constriction injury using Electronic Von Frey. We have been experiencing some challenges with the hand-held electronic Von Frey and animal enclo… --- ## [Experience with electronic von Frey](https://forum.painresearcher.net/t/experience-with-electronic-von-frey/1184)
**Author:** [@cdedek](https://forum.painresearcher.net/u/cdedek)\ **Replies:** 1\ **Last updated:** [March 29, 2024, 4:34am UTC](https://forum.painresearcher.net/t/experience-with-electronic-von-frey/1184 "2024-03-29T04:34:36Z")
I was wondering what peoples’ experience has been with electronic von Frey (eVF). It has been around for a long time, but everyone seems to keep using handheld filaments. I’ve heard eVF is inconsistent, which I can imagi… --- ## [Experience with IB4-Saporin?](https://forum.painresearcher.net/t/experience-with-ib4-saporin/1174)
**Author:** [@PatriciaT](https://forum.painresearcher.net/u/PatriciaT)\ **Replies:** 5\ **Last updated:** [February 1, 2024, 7:44pm UTC](https://forum.painresearcher.net/t/experience-with-ib4-saporin/1174 "2024-02-01T19:44:36Z")
Hi, Anyone has experience using IB4-Saporin from ATS to ablate non peptidergic afferents? We’ve started a set of experiments with this using non conjugated saporin as control. Surprisingly, some animals from both groups… --- ## [DRG neuron adherence issue in culture](https://forum.painresearcher.net/t/drg-neuron-adherence-issue-in-culture/1171)
**Author:** [@sshiers](https://forum.painresearcher.net/u/sshiers)\ **Replies:** 2\ **Last updated:** [January 27, 2024, 3:57pm UTC](https://forum.painresearcher.net/t/drg-neuron-adherence-issue-in-culture/1171 "2024-01-27T15:57:21Z")
Hi all, We are getting some really great neuronal cultures, but once we begin the fixation and ICC processing, the neurons come off the coverslip. We are fixing with cold 10% formalin, and using 1X PBS for washes. The … --- ## [DiI labeling and fixation](https://forum.painresearcher.net/t/dii-labeling-and-fixation/982)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 13\ **Last updated:** [December 22, 2023, 8:26pm UTC](https://forum.painresearcher.net/t/dii-labeling-and-fixation/982 "2023-12-22T20:26:55Z")
I’ve been trying to retrogradely label DRG neurons from various sites. All the labeling methods have their advantages and drawbacks. DiI is commonly used. I have a 5 mg/ml stock in DMSO. I’ve noticed that it’s kind o… --- ## [Precellys 24 for homogenization of mouse dorsal root ganglia](https://forum.painresearcher.net/t/precellys-24-for-homogenization-of-mouse-dorsal-root-ganglia/1047)
**Author:** [@achamess](https://forum.painresearcher.net/u/achamess)\ **Replies:** 7\ **Last updated:** [December 20, 2023, 1:41pm UTC](https://forum.painresearcher.net/t/precellys-24-for-homogenization-of-mouse-dorsal-root-ganglia/1047 "2023-12-20T13:41:54Z")
Does anyone use the Precellys 24 for homogenization of tissues to make protein lysates from mouse DRG or other neural tissues? We have it. I’ve been using it and comparing to mortar-pestle (as @tberta taught me way back… --- ## [Whole mount DRG staining](https://forum.painresearcher.net/t/whole-mount-drg-staining/1131)
**Author:** [@jmajor](https://forum.painresearcher.net/u/jmajor)\ **Replies:** 10\ **Last updated:** [December 13, 2023, 7:43pm UTC](https://forum.painresearcher.net/t/whole-mount-drg-staining/1131 "2023-12-13T19:43:42Z")
Hi everyone, I’m new to pain research and would like to try whole mount stain for murine DRG tissue. I’m wondering, does the DRG require tissue clearing? If so, any tips for how to physically handle a tissue-cleared DRG… --- ## [In vitro Model of CFA-induced inflammatory pain-like response](https://forum.painresearcher.net/t/in-vitro-model-of-cfa-induced-inflammatory-pain-like-response/1155)
**Author:** [@vhms0706](https://forum.painresearcher.net/u/vhms0706)\ **Replies:** 3\ **Last updated:** [December 12, 2023, 9:03am UTC](https://forum.painresearcher.net/t/in-vitro-model-of-cfa-induced-inflammatory-pain-like-response/1155 "2023-12-12T09:03:11Z")
Hello! Does anybody know what would be a good in vitro model (in the context of primary DRG culture calcium imaging experiments) of CFA-induced inflammatory pain behavioral model? I have seen some literature doing the … [Next page](https://forum.painresearcher.net/c/experimental-techniques/6.md?page=1)