Name: Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome (Video) | JoVE Journal
Uploaded: 2016-03-04T13:00:00.000+00:00
Duration: 7 min 16 s
Description: Learn a safe muscle biopsy technique for the vastus lateralis using the Weil-Blakesley conchotome. This method obtains sufficient tissue for histochemistry and molecular analysis with minimal participant discomfort.

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5 minutes

We wish to thank the study participants for making this work possible and the staff at the NIHR Southampton Biomedical Research Centre and the Wellcome Trust Clinical Research Facility, University Hospital Southampton for assistance with study measurements.
This work was supported by the NIHR Southampton Biomedical Research Centre, Nutrition, the MRC Lifecourse Epidemiology Unit and the University of Southampton. This report is independent research by the NIHR BRC Funding Scheme. The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health. AMB is supported by a NIHR Clinical Research Fellowship, HPP is supported by the NIHR Southampton Biomedical Research Centre.

Ethics statement: A description of the muscle biopsy procedure we follow within our institution now follows. The muscle biopsy procedure is used in the Hertfordshire Sarcopenia Study which has been approved by the Hertfordshire Research Committee number 07/Q0204/68. All participants gave written informed consent 20. 
NOTE: A single operator can perform the procedure, aided by an assistant who can engage the participant in conversation in order to reduce anxiety and ensure that they are comfortable at all times. The procedure room should be equipped with a sink, a height adjustable couch, clean surfaces and a procedure trolley.
1. Prepare the Participant (Figure 2)
<ol>
	<li>Ask the participant to lay comfortably, supine on a couch. Expose the preferred thigh from the groin crease. Spread a disposable absorbent sheet under the exposed thigh. Ensure that the leg remains straight and relaxed throughout the procedure but the thigh is tensed momentarily to outline the vastus lateralis.</li>
	<li>Mark the procedure site approximately two-thirds down an imaginary line from the anterior superior iliac spine to the patella.</li>
</ol>
2. Prepare the Skin (Figure 2)
<ol>
	<li>Shave the skin to approximately 4 cm&#160;in diameter surrounding the proposed biopsy site with a sterile shaving blade and clean the skin with an alcohol swab. 
	NOTE: This is to prevent any subsequent hair entrapment in the wound during the healing process. This step is optional according to the operator&#39;s preference.</li>
	<li>Infiltrate the skin and overlying fascia with local anesthetic. Use a 25 G needle to raise a subcutaneous bleb of 2% lidocaine then infiltrate deeper into the subcutaneous tissue with a 23 G needle. Aim to penetrate to at least half to one inch depending on the subcutaneous tissue content at the biopsy site. This depth should be sufficient to ensure the overlying fascia is anesthetized. Allow 2 - 5 min&#160;for the anesthetic to action. Confirm anesthesia by probing the skin gently with the needle or scalpel blade tip.</li>
</ol>
3. Prepare a Sterile Field (Figure 2)
<ol>
	<li>Wash hands with soap and maintain clean hands until a sterile gown is worn. Cover hands with sterile gloves before creating a sterile field on the procedure trolley.</li>
	<li>Sterilize the biopsy site with 2% chlorhexidine/isopropyl alcohol solution or an iodine based skin disinfectant. Apply a sterile drape with an adhesive aperture to expose the biopsy site but to also maintain a sterile field.</li>
</ol>
4. Perform the Procedure (Figure 2)
<ol>
	<li>With a size 11 scalpel, make a 5 - 10 mm incision on the skin and down and through the overlying fascia. Insert the closed biting tip of the conchotome through the incision at a right angle to the long axis of the femur, to a depth averaging half to one inch. If required use the free hand to hold and support the thigh surrounding the biopsy site while the tip of the conchotome is opened and closed within the muscle.</li>
	<li>Rotate the conchotome through 90 - 180&#176; to cut the muscle. Withdraw the conchotome and open the tip onto sterile gauze dampened by sterile normal saline. Repeat the procedure within the single wound site, if necessary, to obtain sufficient muscle tissue. Transfer the gauze into a container placed on ice. Transfer the container to a preparation laboratory to process the muscle according to the local institutional protocol.</li>
</ol>
5. Post Procedure (Figure 2)
<ol>
	<li>Apply direct pressure to the wound for up to 5 min. Close the wound with steri-strips by placing them parallel to the wound as opposed to right angles to avoid potentially removing scar tissue and reopening the wound if the steri-strips are removed inadvertently. Place a sterile absorbent dressing on the steri-strips and tie a two-layer bandage for compression and secure with tape.</li>
	<li>Explain the wound dressing method to the participant as they will have to perform a dressing change 3 - 4 days after the procedure. Ensure that the participant removes the compression bandage before going to bed later that evening.</li>
</ol>
6. Post Biopsy Advice
<ol>
	<li>Inform the participant that it is common to experience some thigh stiffness which can be relieved by gentle exercise (e.g., walking). Use simple analgesia such as acetaminophen. Warn the participant about transient numbness around the biopsy site that may persist for up to two weeks as well as the potential complication of wound infection.</li>
</ol>
7. Instructions for the Subject Post Procedure
<ol>
	<li>Ask the participant to avoid vigorous activity for 72 hr (such as climbing, running, heavy lifting). Ask the participant to avoid immersion in water for 48 - 72 hr. However, instruct the participant to wrap cling film or similar around the biopsy site when having a shower to keep the dressing dry.</li>
	<li>Ask the participant to change the dressing after 3 - 4 days and remove the steri-strips after one week. 
	NOTE: The participant should be given written instructions for post biopsy care, spare dressings and emergency contact details of the team responsible for performing the procedure.</li>
</ol>

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<li>Parent, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Duchenne+De+Boulogne%3A+a+pioneer+in+neurology+and+medical+photography%5BTitle%5D)+AND+%22Can+J+Neurol+Sci%22%5BJournal%5D)">Duchenne De Boulogne: a pioneer in neurology and medical photography.</a> Can J Neurol Sci. 32 (3), 369-377 (2005).</li>
<li>Hayot, M., Michaud, A., Koechlin, C., Caron, M. A., Leblanc, P., Prefaut, C., Maltais, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+muscle+microbiopsy%3A+a+validation+study+of+a+minimally+invasive+technique%5BTitle%5D)+AND+%22Eur+Respir+J%22%5BJournal%5D)">Skeletal muscle microbiopsy: a validation study of a minimally invasive technique.</a> Eur Respir J. 25 (3), 431-440 (2005).</li>
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<li>O'Rourke, K. S., Ike, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+biopsy%5BTitle%5D)+AND+%22Curr+Opin+Rheumatol%22%5BJournal%5D)">Muscle biopsy.</a> Curr Opin Rheumatol. 7 (6), 462-468 (1995).</li>
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<li>Edwards, R., Young, A., Wiles, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Needle+biopsy+of+skeletal+muscle+in+the+diagnosis+of+myopathy+and+the+clinical+study+of+muscle+function+and+repair%5BTitle%5D)+AND+%22N+Engl+J+Med%22%5BJournal%5D)">Needle biopsy of skeletal muscle in the diagnosis of myopathy and the clinical study of muscle function and repair.</a> N Engl J Med. 302 (5), 261-271 (1980).</li>
<li>Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Percutaneous+needle-biopsy+of+skeletal+muscle+in+diagnosis+and+research%5BTitle%5D)+AND+%22Lancet%22%5BJournal%5D)">Percutaneous needle-biopsy of skeletal muscle in diagnosis and research.</a> Lancet. 2 (7724), 593-595 (1971).</li>
<li>Edwards, R. H., Round, J. M., Jones, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Needle+biopsy+of+skeletal+muscle%3A+a+review+of+10+years+experience%5BTitle%5D)+AND+%22Muscle+Nerve%22%5BJournal%5D)">Needle biopsy of skeletal muscle: a review of 10 years experience.</a> Muscle Nerve. 6 (9), 676-683 (1983).</li>
<li>Shanely, R. A., Zwetsloot, K. A., Triplett, N. T., Meaney, M. P., Farris, G. E., Nieman, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Human+skeletal+muscle+biopsy+procedures+using+the+modified+Bergstrom+technique%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Human skeletal muscle biopsy procedures using the modified Bergstrom technique.</a> J Vis Exp. (91), e51812(2014).</li>
<li>Coggan, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+biopsy+as+a+tool+in+the+study+of+aging%5BTitle%5D)+AND+%22J+Gerontol+A+Biol+Sci+Med+Sci%22%5BJournal%5D)">Muscle biopsy as a tool in the study of aging.</a> J Gerontol A Biol Sci Med Sci. 50, 30-34 (1995).</li>
<li>Henriksson, K. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((%27Semi-open%27+muscle+biopsy+technique.+A+simple+outpatient+procedure%5BTitle%5D)+AND+%22Acta+Neurol+Scand%22%5BJournal%5D)">'Semi-open' muscle biopsy technique. A simple outpatient procedure.</a> Acta Neurol Scand. 59 (6), 317-323 (1979).</li>
<li>Dorph, C., Nennesmo, I., Lundberg, I. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Percutaneous+conchotome+muscle+biopsy.+A+useful+diagnostic+and+assessment+tool%5BTitle%5D)+AND+%22J+Rheumatol%22%5BJournal%5D)">Percutaneous conchotome muscle biopsy. A useful diagnostic and assessment tool.</a> J Rheumatol. 28 (7), 1591-1599 (2001).</li>
<li>Poulsen, M. B., Bojsen-Moller, M., Jakobsen, J., Andersen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Percutaneous+conchotome+biopsy+of+the+deltoid+and+quadricep+muscles+in+the+diagnosis+of+neuromuscular+disorders%5BTitle%5D)+AND+%22J+Clin+Neuromuscul+Dis%22%5BJournal%5D)">Percutaneous conchotome biopsy of the deltoid and quadricep muscles in the diagnosis of neuromuscular disorders.</a> J Clin Neuromuscul Dis. 7 (1), 36-41 (2005).</li>
<li>Dietrichson, P., Mellgren, S. I., Skre, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+biopsy+with+the+percutaneous+conchotome+technique%5BTitle%5D)+AND+%22J+Oslo+City+Hosp%22%5BJournal%5D)">Muscle biopsy with the percutaneous conchotome technique.</a> J Oslo City Hosp. 30 (5), 73-79 (1980).</li>
<li>Dietrichson, P., Coakley, J., Smith, P. E., Griffiths, R. D., Helliwell, T. R., Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Conchotome+and+needle+percutaneous+biopsy+of+skeletal+muscle%5BTitle%5D)+AND+%22J+Neurol+Neurosurg+Psychiatry%22%5BJournal%5D)">Conchotome and needle percutaneous biopsy of skeletal muscle.</a> J Neurol Neurosurg Psychiatry. 50 (11), 1461-1467 (1987).</li>
<li>Andonopoulos, A. P., Papadimitriou, C., Melachrinou, M., Meimaris, N., Vlahanastasi, C., Bounas, A., Georgiou, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Asymptomatic+gastrocnemius+muscle+biopsy%3A+an+extremely+sensitive+and+specific+test+in+the+pathologic+confirmation+of+sarcoidosis+presenting+with+hilar+adenopathy%5BTitle%5D)+AND+%22Clin+Exp+Rheumatol%22%5BJournal%5D)">Asymptomatic gastrocnemius muscle biopsy: an extremely sensitive and specific test in the pathologic confirmation of sarcoidosis presenting with hilar adenopathy.</a> Clin Exp Rheumatol. 19 (5), 569-572 (2001).</li>
<li>Helliwell, T. R., Coakley, J., Smith, P. E., Edwards, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+morphology+and+morphometry+of+the+normal+human+tibialis+anterior+muscle%5BTitle%5D)+AND+%22Neuropathol+Appl+Neurobiol%22%5BJournal%5D)">The morphology and morphometry of the normal human tibialis anterior muscle.</a> Neuropathol Appl Neurobiol. 13 (4), 297-307 (1987).</li>
<li>Patel, H. P., Jameson, K. A., Syddall, H. E., Martin, H. J., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Developmental+influences%2C+muscle+morphology%2C+and+sarcopenia+in+community-dwelling+older+men%5BTitle%5D)+AND+%22J+Gerontol+A+Biol+Sci+Med+Sci%22%5BJournal%5D)">Developmental influences, muscle morphology, and sarcopenia in community-dwelling older men.</a> J Gerontol A Biol Sci Med Sci. 67 (1), 82-87 (2012).</li>
<li>Patel, H. P., Syddall, H. E., Martin, H. J., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hertfordshire+Sarcopenia+Study%3A+design+and+methods%5BTitle%5D)+AND+%22BMC+Geriatrics%22%5BJournal%5D)">Hertfordshire Sarcopenia Study: design and methods.</a> BMC Geriatrics. 10 (1), 43(2010).</li>
<li>Anderson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recommendations+for+the+biopsy+procedure+and+assessment+of+skeletal+muscle+biopsies%5BTitle%5D)+AND+%22Virchows+Arch%22%5BJournal%5D)">Recommendations for the biopsy procedure and assessment of skeletal muscle biopsies.</a> Virchows Arch. 431 (4), 227-233 (1997).</li>
<li>Patel, H. P., Syddall, H. E., Martin, H. J., Cooper, C., Stewart, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+Feasibility+and+Acceptability+of+Muscle+Biopsy+in+Epidemiological+Studies%3A+Findings+from+the+Hertfordshire+Sarcopenia+Study+%28HSS%29%5BTitle%5D)+AND+%22J+Nutr+Health+Aging%22%5BJournal%5D)">The Feasibility and Acceptability of Muscle Biopsy in Epidemiological Studies: Findings from the Hertfordshire Sarcopenia Study (HSS).</a> J Nutr Health Aging. 15 (1), 10-15 (2011).</li>
<li>Charifi, N., Kadi, F., Feasson, L., Costes, F., Geyssant, A., Denis, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Enhancement+of+microvessel+tortuosity+in+the+vastus+lateralis+muscle+of+old+men+in+response+to+endurance+training%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Enhancement of microvessel tortuosity in the vastus lateralis muscle of old men in response to endurance training.</a> J Physiol. 554 (Pt 2), 559-569 (2004).</li>
<li>Frontera, W. R., Suh, D., Krivickas, L. S., Hughes, V. A., Goldstein, R., Roubenoff, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+muscle+fiber+quality+in+older+men+and+women%5BTitle%5D)+AND+%22Am+J+Physiol+Cell+Physiol%22%5BJournal%5D)">Skeletal muscle fiber quality in older men and women.</a> Am J Physiol Cell Physiol. 279 (3), C611-C618 (2000).</li>
<li>He, J., Watkins, S., Kelley, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+muscle+lipid+content+and+oxidative+enzyme+activity+in+relation+to+muscle+fiber+type+in+type+2+diabetes+and+obesity%5BTitle%5D)+AND+%22Diabetes%22%5BJournal%5D)">Skeletal muscle lipid content and oxidative enzyme activity in relation to muscle fiber type in type 2 diabetes and obesity.</a> Diabetes. 50 (4), 817-823 (2001).</li>
<li>Patel, H. P., Al-Shanti, N., Davies, L. C., Barton, S. J., Grounds, M. D., Tellam, R. L., Stewart, C. E., Cooper, C., Sayer, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Lean+mass%2C+muscle+strength+and+gene+expression+in+community+dwelling+older+men%3A+findings+from+the+Hertfordshire+Sarcopenia+Study+%28HSS%29%5BTitle%5D)+AND+%22Calcif+Tissue+Int%22%5BJournal%5D)">Lean mass, muscle strength and gene expression in community dwelling older men: findings from the Hertfordshire Sarcopenia Study (HSS).</a> Calcif Tissue Int. 95 (4), 308-316 (2014).</li>
<li>Peterson, C. M., Johannsen, D. L., Ravussin, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+muscle+mitochondria+and+aging%3A+a+review%5BTitle%5D)+AND+%22J+Aging+Res%22%5BJournal%5D)">Skeletal muscle mitochondria and aging: a review.</a> J Aging Res. 2012, 194821(2012).</li>
</ol>

None declared.

To ensure participant safety and perform the muscle biopsy efficiently, it is vital to pay attention to critical steps within the protocol.
The participants must be selected after carefully considering the exclusion criteria to avoid complications such as bleeding and poor wound healing. Strict asepsis throughout the procedure is essential. Application of the correct technique will ensure participant comfort, minimize unwanted trauma to participant&#39;s tissues and allow a sufficient sample y...

Obtaining skeletal muscle for the diagnosis of a myopathy and other neuromuscular degenerative diseases relies on safe and efficient methods which are acceptable and not incapacitating to patients 1. Historically, methods of obtaining muscle tissue for analysis included open biopsies performed under general anesthesia or from post mortem. These techniques allowed direct visualization of the muscle and permitted a larger piece of muscle to be biopsied. Given the potential disadvantages these techniques have to the patient in terms of hospital stay and recovery, percutaneous &#39;semi-open&#39; muscle biopsy methods were introduced as alternatives. This technique was first performed by Duchenne (1806-1875), who used a self-constructed needle with a trocar and was able to obtain a muscle sample from a living subject without general anesthesia 2. Since then, various forms of biopsy needles have been used 3,4,5 . However, the biopsy needle developed by Bergstrom in 1962 has been the most commonly used in clinical practice; both in children and adults, as well as in research 6,7,8,9,10. It possesses a sharp trocar, a cutting cannula and a pushing rod to expel the tissue post biopsy. Muscle yields obtained from this procedure have been reported to range from 25 -293 mg after repeated sampling 11,9.
Henriksson introduced the Weil-Blakesley conchotome in 1979 as an alternative semi-open muscle biopsy method (Figure 1) 12. The instrument is a single structure unlike the Bergstrom construct and is designed like a forcep with a sharp biting tip. The tip size can range from 4 mm to 6 mm in width. When the two edges of the biting tip oppose, a hollow is formed that ensures capture of the muscle. The conchotome is inserted through a 5 - 10 mm skin incision and avoids the need for a trocar to penetrate the muscle and the overlying fascia as a scalpel can be used to make a track down to the muscle.
This method allows controlled tissue penetration with a high degree of maneuverability 13,14 . It is especially useful for sites where undue pressure could potentially damage neurovascular or underlying bony structures e.g., at the site of the tibialis anterior 15,14. Muscle yields using the conchotome can be variable and in our practice 20 - 200 mg of muscle has been obtained after repeated sampling. One potential advantage of the conchotome over the Bergstrom needle, in addition to the high degree of maneuverability it offers, is that the biting tip does not need to be sharpened or replaced as often as the trocar of the Bergstrom construct 16 .
The overall goal of muscle biopsy using the Weil Blakesley conchotome is to obtain sufficient amounts of muscle tissue to enable histochemistry and molecular analysis for both diagnostic and research study. It is a simple and safe procedure that can be learned easily. With this technique, muscle suitable for biopsy includes biceps, triceps, deltoid, gastrocnemius, tibialis anterior, soleus and the sacrospinals 17,15,7,18,14 . The outermost part of the vastus lateralis is the most common site used for biopsy as it avoids the main neurovascular structures within the thigh. It is identified approximately two thirds down an imaginary line joining the anterior superior iliac spine and the patella. Muscle morphology data, for example myofibre area, myofibre proportions, capillary density derived from vastus lateralis sampling are widely available in the literature allowing comparison between studies 11,19 .

<table><tbody><tr><td>Weil-Blakesley conchotome</td><td>Gebr&amp;uuml;der Zepf Medizintechnik, D&amp;uuml;rbheim, Germany</td><td>None</td><td>6 mm biting tip</td></tr><tr><td>5 ml 2% lidocaine</td><td>Generic use</td><td>None</td><td>One 5 ml vial should suffice for adequate anaesthesia</td></tr><tr><td>Alcohol swab</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>5 ml syringe</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>25 G and 23 G sterile needles</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile scalpel size 11</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gauze squares</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sodium chloride 0.9% 10 ml</td><td>Fannin</td><td>1122/002/001</td><td></td></tr><tr><td>Steri-Drape&amp;trade; Small Drape with Adhesive Aperture</td><td>3M</td><td>1020</td><td></td></tr><tr><td>ChloraPrep 2% w/v /70% v/v cutaneous solution&amp;nbsp;</td><td>CareFusion</td><td>270400</td><td>Two</td></tr><tr><td>Steri-Strip&amp;trade; 1/4 inch x 3 inch Reinforced Adhesive Skin Closures</td><td>3M</td><td>R1541</td><td></td></tr><tr><td>PRIMAPORE sterile dressing 15 cm x 8 cm</td><td>Smith &amp;amp;&amp;nbsp;&lt;br/&gt; Nephew</td><td>66000318</td><td></td></tr><tr><td>Cotton crepe bandage Hospicrepe 233 7,5 cm x 4,5 m</td><td>Hartmann</td><td>915562</td><td></td></tr><tr><td>Flexible cohesive bandage Co-Plus LF</td><td>BSN medical</td><td>2904565</td><td></td></tr><tr><td>Disposable absorbent sheet</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Micropore or transpore tape</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gown with sterile paper towels</td><td>Generic use</td><td>None</td><td></td></tr><tr><td>Sterile gloves</td><td>Generic use</td><td>None</td><td></td></tr></tbody></table>

human vastus lateralis skeletal muscle biopsy using the weil-blakesley conchotome

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and well tolerated technique. The Weil-Blakesley conchotome has a sharp biting tip with a 4 - 6 mm wide hollow. It is inserted through a 5 - 10 mm skin incision and can be maneuvered for controlled tissue penetration. The tip is opened and closed within the tissue and then rotated through 90 -180&#176; to cut the muscle. The amount of muscle obtained following repeated sampling can vary from 20 mg to 290 mg which can be processed for both histology and molecular studies. The wound needs to be kept dry and vigorous physical activity kept to a minimum for approximately 72 hr although normal levels of activity can restart immediately following the procedure. This procedure is safe and effective when close attention is paid to the selection of subjects, full asepsis and post procedure care. &#160;Both right and left vastus lateralis are suitable for biopsy dependent on participant preference.

The procedure described above is safe and acceptable in both clinical and research settings. When the biopsy is performed as part of the diagnostic process, the muscle should be chosen according to patient's symptoms and signs of muscular weakness. The biopsy site should be free from previous injuries, contractures or instrumentation 21,1. In research, standardized conditions such as fasting or exclusion of patients with diabetes may be required 19. Anticoagulants su...

This video demonstrates the technique of percutaneous muscle biopsy of the human vastus lateralis using the Weil-Blakesley conchotome.

Percutaneous muscle biopsy using the Weil-Blakesley conchotome is well established in both clinical and research practice. It is a safe, effective and ...

The overall goal of this method is to introduce a safe, effective, and well-tolerated technique of muscle biopsy of the vastus lateralis, to obtain sufficient tissue for histochemistry and molecular analysis in both diagnostic and research studies. This method can help investigators obtain sufficient muscle tissue with minimal discomfort and trauma for the participant. The main advantage of this technique is that it's safe and acceptable in both clinical and research settings.Complications are low, and discomfort's kept to a minimum when the procedure is performed using a standard technique. To begin, ask the participant to lie comfortably on a couch in a supine position. Expose the preferred thigh from the groin crease, and spread a disposable absorbent sheet under the exposed thigh.Ensure that the leg remains straight and relaxed throughout the procedure, but the thigh is tensed momentarily to outline the vastus lateralis. Mark the procedure site approximately 2/3 down an imaginary line from the anterior superior iliac spine to the patella. To prepare the skin, use a sterile shaving blade to shave approximately four centimeters in diameter surrounding the proposed biopsy site, in men, if required.Use an alcohol swab to clean the skin. Use a local anesthetic to infiltrate the skin and overlying fascia, then, with a 25-gauge needle, raise a subcutaneous bleb of 2%lidocaine. Next, use a 23-gauge needle to infiltrate deeper into the subcutaneous tissue, aiming to penetrate at least 1/2 an inch, depending on the subcutaneous content at the biopsy site.Allow two to five minutes for the anesthesia to take action, and confirm anesthesia by using a needle or scalpel blade tip to gently probe the skin. To prepare a sterile field, wash hands with soap and water, and maintain clean hands until a sterile gown is worn. Cover hands with sterile gloves before creating a sterile field on the procedure trolley.With 2%Chlorhexidine isopropyl alcohol solution, or an iodine-based skin disinfectant, sterilize the biopsy site. Apply a sterile drape with an adhesive aperture to expose the biopsy site and to maintain a sterile field. The first critical step is to puncture the skin and make a sufficiently deep track into and beyond the fascia.To perform the biopsy procedure, with a size 11 scalpel, make a five to 10 millimeter incision on the skin, and in through the overlying fascia. While holding and supporting the thigh surrounding the biopsy site, insert the closed biting tip of the conchotome through the incision, at a right angle to the long axis of the femur, approximately 1/2 to one inch deep. Next, rotate the conchotome from 90 to 180 degrees to cut the muscle, then withdraw the conchotome and open the tip onto sterile gauze dampened by sterile normal saline.Transfer the gauze into a container placed on ice. The second critical step is to insert the conchotome within the track, to maneuver the conchotome to ensure sufficient muscle has been obtained. If necessary, the procedure can be repeated within the track, and single wound site to obtain sufficient muscle tissue.Following the procedure, apply direct pressure to the wound for up to five minutes. Then place sterile strips parallel to the wound to close it. Place a sterile absorbent dressing on the Steri-Strips, and apply a two-layer bandage for compression, using tape to secure.Explain the wound dressing method to the participant for them to perform three to four days after the procedure. Instruct the participant to remove the compression bandage before going to bed later that evening. Explain that it is common to experience some thigh stiffness, which can be relieved by gentle exercise, such as walking.Explain that they can use acetaminophen for the pain. As shown in this table, in the Hertfordshire sarcopenia study, the pain score is measured using the pain Visual Analogue Scale of zero millimeters for no pain, to 100 millimeters to pain as bad as can be, were typically low, with a median of seven millimeters during the procedure, and four millimeters one day after the procedure. This suggests that the subjects in this research study found the procedure minimally painful.Shown here are representative tissue samples obtained through percutaneous muscle biopsy with the Weil-Blakesley conchotome. The samples obtained should resemble the muscle tissue in this panel. However, there is a high probability of obtaining subcutaneous fatty tissue, as seen in this panel.This figure shows a serial cross section with differential fiber staining. Darkly stained fibers represent type two fast fibers detected with an anti-myosin fast antibody. Once mastered, this technique can be done in 20 minutes, if it is performed properly.While attempting this procedure, it's important to remember to maintain a sterile field, and keep the participant engaged. Following this procedure, other methods like histochemistry and DNA or RNA extraction can be performed, and all that will answer additional questions, like what are the cellular and molecular changes that occur in aging muscle? After its development, this technique paved way for researchers in the field of muscle biology and aging, to explore the changes in muscle tissue with sarcopenia, the age-related decline of muscle mass and function.After watching this video, you should have a good understanding of how to perform a muscle biopsy of the vastus lateralis, using the Weil-Blakesley conchotome, a simple, effective, and well-tolerated procedure. Don't forget that working with sharp needles and instruments can be hazardous, and standard safety precautions as well as strict asepsis should always be maintained while performing this procedure.

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Research

JoVE Journal

Medicine

The purpose of this video is to present the modified Bergström skeletal muscle biopsy technique on human subjects.

Human Skeletal Muscle Biopsy Procedures Using the Modified Bergstr&#246;m Technique

The main adherent cell types derived from human muscle are myogenic cells and fibroblasts. Here, cell populations are enriched using magnetic-activated cell sorting based on the CD56 antigen. Subsequent immunolabelling with specific antibodies and use of image analysis techniques allows quantification of cytoplasmic and nuclear characteristics in individual cells.

Isolation and Quantitative Immunocytochemical Characterization of Primary Myogenic Cells and Fibroblasts from Human Skeletal Muscle

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

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Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation

3D ultrasound imaging (3DUS) allows fast and cost-effective morphometry of musculoskeletal tissues. We present a protocol to measure muscle volume and fascicle length using 3DUS.

3D Ultrasound Imaging: Fast and Cost-effective Morphometry of Musculoskeletal Tissue

Immunohistochemical staining of myosin heavy chain isoforms has emerged as the state-of-the-art discriminator of skeletal muscle fiber-type (i.e., type I, type IIA, type IIX, type IIB). Here, we present a staining protocol along with a novel semi-automated algorithm that facilitates rapid assessment of fiber-type and fiber morphology.

Semi-automated Analysis of Mouse Skeletal Muscle Morphology and Fiber-type Composition

We present detailed protocols for performing small-angle X-ray diffraction experiments using intact mouse skeletal muscles. With the wide availability of transgenic mouse models for human diseases, this experimental platform can form a useful test bed for elucidating the structural basis of genetic muscle...

X-ray Diffraction of Intact Murine Skeletal Muscle as a Tool for Studying the Structural Basis of Muscle Disease

We present techniques for isolating, culturing, characterizing, and differentiating human primary muscle progenitor cells (hMPCs) obtained from skeletal muscle biopsy tissue. hMPCs obtained and characterized through these methods can be used to subsequently address research questions related to human myogenesis and skeletal muscle regeneration.

Isolation, Culture, Characterization, and Differentiation of Human Muscle Progenitor Cells from the Skeletal Muscle Biopsy Procedure

Described here is a protocol for direct immunolabelling of necrotic myofibers in muscle cryosections. Necrotic cells are permeable to serum proteins, including immunoglobulin G (IgG). Revealing the uptake of IgG by myofibers allows the identification and quantification of myofibers that undergo necrosis regardless of muscle condition.

Immunolabelling Myofiber Degeneration in Muscle Biopsies

This technical report describes a variation of the modified Bergström technique for the biopsy of the musculus tibialis anterior that limits fiber damage.

Collection of Skeletal Muscle Biopsies from the Superior Compartment of Human Musculus Tibialis Anterior for Mechanical Evaluation

This protocol aims to isolate and purify skeletal muscle interstitial extracellular vesicles (SkM-EVs) from rodent muscle tissues through mechanical detachment, enzymatic dissociation, filtration, and differential ultracentrifugation. It demonstrates consistency and reliability. The isolated SkM-EVs provide insights into muscle homeostasis and diseases, with potential applications as diagnostic biomarkers or therapeutic vehicles.

Enzymatic Isolation of Skeletal Muscle Interstitial Extracellular Vesicles

Learn a safe muscle biopsy technique for the vastus lateralis using the Weil-Blakesley conchotome. This method obtains sufficient tissue for histochemistry and molecular analysis with minimal participant discomfort.

Human Vastus Lateralis Skeletal Muscle Biopsy Using the Weil-Blakesley Conchotome

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