אין מימון להכריז.

<ol>
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The authors have no conflicts of interest.

וידאו זה מספק סיכום צעד אחר צעד של הליך ביופסיה של שריר השלד המשמש במעבדה ביצועי אדם ASU. הליך זה, עם שינויים קטנים, נעשה שימוש כדי לאסוף כ 1,600 ביופסיות שריר במהלך 13 השנים האחרונות. דגימות הביופסיה של השריר סיפקו נתונים חשובים בחקירות מבוססות תזונת ספורט, שהובילו לתגליו?...

טכניקת הביופסיה מלעורית או &#39;פתוחה למחצה &quot;משמשת כדי להשיג דגימות רקמת שריר שלד מחולים אנושיים ונושאי מחקר. דושן (1806-1875) הוא זוכה כראשון לבניית מחט עם נקז כדי להשיג שרירי שלד מנושאים המתגוררים דרך הביופסיה מלעורית 1. בשנתי ה -1960, Bergstrom הציג מחט ביופסיה מלעורית דומה לזה שתואר על ידי דושן 2-4. עשרים שנה מאוחר יותר, אוונס et al. 5 הותאם על ידי יישום טכניקת יניקה דרך נקז החיתוך של מחט Bergstrom. שינוי זה יכול לשפר את התשואה של רקמה 3 עד 5 פי 6,7, והוא מועסק בהגדרות מחקר קליניות וביו. טכניקה זו יש ותמשיך לטפח את האבחנה של myopathies וההבנה של המבנה והתפקוד של שרירי שלד שלנו. טכניקת הביופסיה של שריר מלעורית היא straightforward. כאשר נעשו בצורה נכונה, עם הקפדה על טכניקת ספטית, הסיכונים הכרוכים בכך הם מינימאליים. הליך הביופסיה של השריר נחשב ככלי מחקרי לפרויקטי מחקר. חלק מהמוסדות האקדמיים מאפשרים מאומנים חוקרי פקולטה עם תואר דוקטור, עם פיקוח רופא ישיר, כדי להשיג ביופסיות שריר ואחרים דורשים רופא מוסמך לוח כדי לבצע את הטכניקה. צוות הביופסיה ASU ביצע בהצלחה מעל 1,600 ביופסיות שריר במהלך 13 השנים האחרונות 8-15. המטרה של הווידאו הזה היא לתאר את טכניקת הביופסיה של שריר מלעורית מחט Bergstrom הותאם 5 להשיג דגימות רקמת שריר שלד מlateralis vastus של בני אדם.

<table border="0" cellpadding="0" cellspacing="0" width="969">
	<tbody>
		<tr height="24">
			<td height="24" style="height:24px;width:439px;">Name of Material / Equipment</td>
			<td style="width:185px;">Company</td>
			<td style="width:141px;">Catalog Number</td>
			<td style="width:204px;">Comments /&#160; Description</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Bergstr&#246;m biopsy needle, 5 x 100mm</td>
			<td style="width:185px;">Stille Surgical Inc.</td>
			<td>119-29187-50</td>
			<td>1 ea</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Durasorb blue pad</td>
			<td>Fisher</td>
			<td>22-031-340</td>
			<td>case of 300</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Prep Razor Disposable</td>
			<td>Moore Medical</td>
			<td>89760</td>
			<td>1 ea</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Shave Cream</td>
			<td>Moore Medical</td>
			<td>92869</td>
			<td>1 can</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Alcohol Swab</td>
			<td>Moore Medical</td>
			<td>98721</td>
			<td>box/200</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Povidone-iodine Swabsticks (3 pack)</td>
			<td>Moore Medical</td>
			<td>90691</td>
			<td>pack of 50</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Face masks</td>
			<td>Fisher</td>
			<td>19-039-690</td>
			<td>case of 50</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Sterile Gloves; sz 6.5</td>
			<td>Moore Medical</td>
			<td>68128</td>
			<td>box of 50 pairs</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Sterile Gloves; sz 7.0</td>
			<td>Moore Medical</td>
			<td>68129</td>
			<td>box of 50 pairs</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Sterile Gloves; sz 8.5</td>
			<td>Moore Medical</td>
			<td>68132</td>
			<td>box of 50 pairs</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Fenestrated Towel Drape</td>
			<td>Moore Medical</td>
			<td>92712</td>
			<td>case of 50</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Lidocaine (1% HCl, w/o Epi</td>
			<td>Dealmed</td>
			<td>427902</td>
			<td>30cc vial</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Ethyl Chloride spray</td>
			<td>Dealmed</td>
			<td>386020</td>
			<td>3.5 fl. Oz. bottle</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">5 mL syringe w/ 21G needle</td>
			<td>Fisher</td>
			<td>14-827-48</td>
			<td>case of 100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">25G 5/8&#34; needle</td>
			<td>Fisher</td>
			<td>14-826AA</td>
			<td>case of 100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">25G 1 1/2&#34; needle</td>
			<td>Fisher</td>
			<td>14-826-49</td>
			<td>case of 100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Single-Use Scalpels&#160; w/ #11 blade</td>
			<td>Fisher</td>
			<td>0-8927-5B</td>
			<td>box of 20</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">60 mL Syringe</td>
			<td>Fisher</td>
			<td>13-689-8</td>
			<td>case of 40</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">~30 inch extension tubing</td>
			<td>Fisher</td>
			<td>14-169-7A</td>
			<td>roll of 50ft</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Metal stopcock</td>
			<td>Fisher</td>
			<td>01-290-38</td>
			<td>each</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Polypropylene tubing connector w/ tapered ends, 3.2 to 5.5 mm</td>
			<td>Fisher</td>
			<td>15-315-28A</td>
			<td>case of 100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Yellow pipette tips (1-200 &#181;L)</td>
			<td>Fisher</td>
			<td>02-681-2</td>
			<td>cut off&#160; ~15-18 mm of the tip</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Topical surgical adhesive</td>
			<td>Medline</td>
			<td>MSC091076</td>
			<td>case of 36 tubes</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Topical surgical adhesive</td>
			<td>Medline</td>
			<td>MSC091076Z</td>
			<td>box of 6 tubes</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Butterfly stitches (adhesive strips) 1/2&#34; x 4&#34; (6 per pack)</td>
			<td>Moore Medical</td>
			<td>93050</td>
			<td>box/300</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Bandadge, Strip, 3/4 x 3 in.</td>
			<td>Fisher</td>
			<td>19-027-202</td>
			<td>pack of 100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Adhesive bandage (5m rolls)</td>
			<td>Fisher</td>
			<td>19-027-768</td>
			<td>case of 36</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">1&#34; paper surgical tape</td>
			<td>Fisher</td>
			<td>19-027-761</td>
			<td>case of 12</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Sutures (4-0 reverse cutting, FS2)</td>
			<td>Moore Medical</td>
			<td>82926</td>
			<td>box of 36</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">4 x 4 sterile gauze pads</td>
			<td>Moore Medical</td>
			<td>66949</td>
			<td>pkg/50</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">2 x 2 non-sterile gauze pads</td>
			<td>Moore Medical</td>
			<td>37336</td>
			<td>pkg/200</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">3 x 4 Non-Adherent Pads</td>
			<td>Moore Medical</td>
			<td>74389</td>
			<td>bx/100</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Autoclave bags 3.5 x 9</td>
			<td>Moore Medical</td>
			<td>40390</td>
			<td>box of 200</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Autoclave bags 5.25 x 10</td>
			<td>Moore Medical</td>
			<td>40391</td>
			<td>box of 200</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Autoclave indicator tape</td>
			<td>Moore Medical</td>
			<td>31608</td>
			<td>box of 250</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Lab Coats&#8212;small</td>
			<td>Fisher</td>
			<td>23-900-520A&#160;</td>
			<td>10/pk</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">Lab Coats&#8212;medium</td>
			<td>Fisher</td>
			<td>23-900-520B&#160;</td>
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human skeletal muscle biopsy procedures using the modified bergstr&#246;m technique

טכניקת הביופסיה מלעורית מאפשרת לחוקרים ולרופאים כדי לאסוף דגימות רקמת שריר שלד. הטכניקה היא בטוחה ויעילה ביותר. וידאו זה מתאר את טכניקת הביופסיה מלעורית באמצעות מחט Bergstrom שונה כדי להשיג דגימות רקמת שריר שלד מlateralis vastus של בני אדם. מחט Bergstrom מורכבת מצינורית חיצונית עם פתח קטן (&#39;חלון&#39;) בצד של הקצה ונקז פנימי עם להב חיתוך בקצה הדיסטלי. בתנאי הרדמה וספטית מקומיים, המחט התקדמה לתוך שריר השלד באמצעות חתך בעור, הרקמה התת עורית, וfascia. בשלב הבא, היניקה מוחלת על הנקז הפנימי, הנקז החיצוני משוך לאחור, רקמת שריר שלד הוא נשאבה לתוך החלון של הצינורית החיצונית על ידי היניקה, והנקז הפנימי סגור במהירות, ובכך חיתוך או גזיר דגימת רקמת שריר השלד . המחט מסובבת 90 °60; וקיצוץ נוסף הוא עשה. תהליך זה ניתן לחזור עוד שלוש פעמים. טכניקת חיתוך מרובה זה בדרך כלל מייצרת מדגם של 100-200 מ&quot;ג או יותר בנבדקים בריאים ויכולה להיעשות באופן מיידי לפני, במהלך, ואחרי התקף של פעילות גופנית או התערבות אחרת. בעקבות הלבשה לאחר ביופסיה של אתר החתך, נושאים בדרך כלל לחדש את פעילותם של חיים היום יום באופן מיידי ויכולים להשתתף באופן מלא בפעילות גופנית נמרצת בתוך 48-72 שעה. נושאים צריכים להימנע מפעילות גופני התנגדות כבדה ל48 שעות כדי להפחית את הסיכון של פריצת של השריר דרך החתך בfascia.

הליך הביופסיה של השריר כפי שתואר לעיל מאפשר לחוקר לאסוף במהירות ובאופן עקבי דגימות רקמת שריר שלד. התשואה האופיינית בנבדקים בריאים, אתלטי היא 200 מ&quot;ג או יותר במעבר אחד עם 3-4 סרטונים. ההליך אורך 15-20 דקות, שרובו בילו בהכנה לחתך. במחקרים מבוססי תרגיל, דגימות שריר לעתים קרו...

Watch this Scientific Journal Video about Human Skeletal Muscle Biopsy Procedures Using the Modified BergstrÃ¶m Technique at JoVE.com

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

The purpose of this video is to present the modified Bergstr&#246;m skeletal muscle biopsy technique on human subjects.

נהלי ביופסיה של שריר שלד אנושי באמצעות טכניקת Bergstrom השתנה

The percutaneous biopsy technique enables researchers and clinicians to collect skeletal muscle tissue samples. The technique is safe and highly ...

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Research

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Medicine

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

39.5K Views.  Appalacian State University. The purpose of this video is to present the modified Bergström skeletal muscle biopsy technique on human subjects. 

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

Myo-mechanical Analysis of Isolated Skeletal Muscle

To assess the in vivo effects of therapeutic interventions for the treatment of muscle disease 1,2,3, quantitative methods are needed that measure force generation and fatigability in treated muscle. We describe a detailed approach to evaluating myo-mechanical properties in freshly explanted hindlimb muscle from the mouse. We describe the atraumatic harvest of mouse extensor digitorum longus muscle, mounting the muscle in a muscle strip myograph (Model 820MS; Danish Myo Technology), and the measurement of maximal twitch and tetanic tension, contraction time, and half-relaxation time, using a square pulse stimulator (Model S48; Grass Technologies). Using these measurements, we demonstrate the calculation of specific twitch and tetanic tension normalized to muscle cross-sectional area, the twitch-to-tetanic tension ratio, the force-frequency relationship curve and the low frequency fatigue curve 4. This analysis provides a method for quantitative comparison between therapeutic interventions in mouse models of muscle disease 1,2,3,5, as well as comparison of the effects of genetic modification on muscle function 6,7,8,9.

To assess the in vivo effects of therapeutic interventions for muscle disease, methods are needed to quantitate force generation and fatigability in treated muscle. We detail an approach to evaluating myo-mechanical properties in explanted mouse hindlimb muscle. This analysis provides a robust approach to quantitating the effects of genetic modification on muscle function, as well as comparison of therapies in mouse models of muscle disease.

מיו מכני ניתוח בשרירי השלד מבודדת

To assess the in vivo effects of therapeutic interventions for the treatment of muscle disease 1,2,3, quantitative methods are needed that measure force ...

Skeletal Muscle Gender Dimorphism from Proteomics

Gross contraction in skeletal muscle is primarily determined by a relatively small number of contractile proteins, however this tissue is also remarkably adaptable to environmental factors1 such as hypertrophy by resistance exercise and atrophy by disuse. It thereby exhibits remodeling and adaptations to stressors (heat, ischemia, heavy metals, etc.)2,3. Damage can occur to muscle by a muscle exerting force while lengthening, the so-called eccentric contraction4. The contractile proteins can be damaged in such exertions and need to be repaired, degraded and/or resynthesized; these functions are not part of the contractile proteins, but of other much less abundant proteins in the cell. To determine what subset of proteins is involved in the amelioration of this type of damage, a global proteome must be established prior to exercise5 and then followed subsequent to the exercise to determine the differential protein expression and thereby highlight candidate proteins in the adaptations to damage and its repair. Furthermore, most studies of skeletal muscle have been conducted on the male of the species and hence may not be representative of female muscle. 
In this article we present a method for extracting proteins reproducibly from male and female muscles, and separating them by two-dimensional gel electrophoresis followed by high resolution digital imaging6. This provides a protocol for spots (and subsequently identified proteins) that show a statistically significant (p &lt; 0.05) two-fold increase or decrease, appear or disappear from the control state. These are then excised, digested with trypsin and separated by high-pressure liquid chromatography coupled to a mass spectrometer (LC/MS) for protein identification (LC/MS/MS)5. This methodology (Figure 1) can be used on many tissues with little to no modification (liver, brain, heart etc.).

A straight-forward set of methods to isolate and determine the identity of the most abundant proteins expressed in skeletal muscle. About 800 spots are discerned on a two-dimensional gel from 10 mg muscle; this allows for the determination of gender-specific protein expression. These methods will give equivalent results in most tissues.

שרירי השלד מין Dimorphism מ Proteomics

Gross contraction in skeletal muscle is primarily determined by a relatively small number of contractile proteins, however this tissue is also remarkably ...

Skin Punch Biopsy Explant Culture for Derivation of Primary Human Fibroblasts

Tissues and cell lines derived from an individual with disease are ideal sources to study disease-related cellular phenotypes. Patient-derived fibroblasts in this protocol have been successfully used in the derivation of induced pluripotent stem cells to model disease1. Early passages of these fibroblasts can also be used for cell-based functional assays to study specific disease pathways, mechanisms2 and subsequent drug screening approaches. The advantage of the presented protocol over enzymatic procedures are 1) the reproducibility of the technique from small amounts of tissue derived from older patients, e.g. patients affected with Parkinson's disease, 2) the technically simple approach over more challenging methodologies using enzymatic treatments, and 3) the time consideration: this protocol takes 15-20 min and can be performed immediately after biopsy arrival. Enzymatic treatments can take up to 4 hr and have the problems of overdigestion, reduction of cell viability and subsequent attachment of cells when not handled properly. This protocol describes the dissection and preparation of a 4-mm human skin biopsy for derivation of a fibroblast culture and has a very high success rate which is important when dealing with patient-derived tissue samples. In this culture, keratinocytes migrate out of the biopsy tissue within the first week after preparation. Fibroblasts appear 7-10 days after the first outgrowth of keratinocytes. DMEM high glucose media supplemented with 20% FBS favors the growth of fibroblasts over keratinocytes and fibroblasts will overgrow the keratinocytes. After 2 passages keratinocytes have been diluted out resulting in relatively homogenous fibroblast cultures which expresses the fibroblast marker SERPINH1 (HSP-47). Using this approach, 15-20 million fibroblasts can be derived in 4-8 weeks for cell banking. The skin dissection takes about 15-20 min, cells are then monitored once a day under the microscope, and media is changed every 2-3 days after attachment and outgrowth of cells.

The fibroblast explant culture protocol from human skin punch biopsies is a technically robust and simple way to derive skin cells within 4-8 weeks for banking of about 15-20 million cells at a low passage number.

תרבות explant עור אגרוף ביופסיה לגזירה של fibroblasts אדם היסודי

Tissues and cell lines derived from an individual with disease are ideal sources to study disease-related cellular phenotypes. Patient-derived fibroblasts ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

חיתוך רוחב עצב הטיביאלי - מודל סטנדרטי לניוון שרירים שלד מושרה denervation בעכברים

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are available to perform mechanistic in vivo studies. While heart transplantation models using a suture technique were first successfully developed in rats, the translation into an equally widespread used murine equivalent was never achieved due the technical complexity of the microsurgical procedure. In contrast, non-suture cuff techniques, also developed initially in rats, were successfully adapted for use in mice1-3. This technique for revascularization involves two major steps I) everting the recipient vessel over a polyethylene cuff; II) pulling the donor vessel over the formerly everted recipient vessel and holding it in place with a circumferential tie. This ensures a continuity of the endothelial layer, short operating time and very high patency rates4.
Using this technique for vascular anastomosis we performed more than 1,000 cervical heart transplants with an overall success rate of 95%. For arterial inflow the common carotid artery and the proximal aortic arch were anastomosed resulting in a retrograde perfusion of the transplanted heart. For venous drainage the pulmonary artery of the graft was anastomosed with the external jugular vein of the recipient5.
Herein, we provide additional details of this technique to supplement the video.

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.
Herein, we provide additional details of this technique to supplement the video.

עכברי צוואר הרחם לב השתלת דגם שימוש בטכניקת חפתים השתנה

Mouse models are of special interest in research since a wide variety of monoclonal antibodies and commercially defined inbred and knockout strains are ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

Assay המבוסס תא זרימת cytometry תפוקה גבוהה לזיהוי נוגדנים לN-מתיל-D-aspartate קולטן דופמין או-2 קולטן בסרום אדם

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

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

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

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.

ההכנה וRespirometric הערכה של המיטוכונדריה מבודדת מרקמת שריר שלד מתקבלת על ידי Percutaneous מחט ביופסיה

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

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.

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

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

בן אנוש<em&gt; Vastus lateralis</em&gt; ביופסיה בשרירי השלד שימוש וייל-בלייסלי Conchotome

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

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo and noninvasively in humans via phosphorus-31 magnetic resonance spectroscopy (31PMRS). However, the approach has not been widely adopted in translational and clinical research, with variations in methodology and limited guidance from the literature. Increased optimization, standardization, and dissemination of methods for in vivo 31PMRS would facilitate the development of targeted therapies to improve OXPHOS capacity and could ultimately favorably impact cardiovascular health. 31PMRS produces a noninvasive, in vivo measure of OXPHOS capacity in human skeletal muscle, as opposed to alternative measures obtained from explanted and potentially altered mitochondria via muscle biopsy. It relies upon only modest additional instrumentation beyond what is already in place on magnetic resonance scanners available for clinical and translational research at most institutions. In this work, we outline a method to measure in vivo skeletal muscle OXPHOS. The technique is demonstrated using a 1.5 Tesla whole-body MR scanner equipped with the suitable hardware and software for 31PMRS, and we explain a simple and robust protocol for in-magnet resistive exercise to rapidly fatigue the quadriceps muscle. Reproducibility and feasibility are demonstrated in volunteers as well as subjects over a wide range of functional capacities.

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

This work demonstrates the feasibility of an in vivo phosphorus-31 magnetic resonance spectroscopy (31PMRS) technique to quantify mitochondrial oxidative phosphorylation (OXPHOS) capacity in human skeletal muscle.

זרחן-31 ספקטרוסקופיה תהודה מגנטית: כלי למדידת<em&gt; In vivo</em&gt; קיבולת זרחון חמצוני מיטוכונדריאלי של שריר שלד אדם

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...

Mouse Model for Pancreas Transplantation Using a Modified Cuff Technique

Mouse models have several advantages in transplantation research, including easy handling, a variety of genetically well-defined strains, and the availability of the widest range of molecular probes and reagents to perform in vivo as well as in vitro studies. Based on our experience with various murine transplantation models, we developed a heterotopic pancreas transplantation model in mice with the intent to analyze mechanisms underlying severe ischemia reperfusion injury-associated early graft damage. In contrast to previously described techniques using suture techniques, herein we describe a new procedure using a non-suture cuff technique. 
In recent years, we have performed more than 300 pancreas transplantations in mice with an overall success rate of &gt;90%, a success rate never described before in mouse pancreas transplantation. The backbone of this non-suture cuff technique for graft revascularization consists of two major steps: (I) pulling the recipient vessel over a polyethylene/polyamide cuff and fixing it with a circumferential ligature, and (II) placing the donor vessel over the everted recipient vessel and fixing it with a second circumferential ligature. The resultant continuity of the endothelial layer results in less thrombogenic lesions with high patency rates and, finally, high success rates.
In this model, arterial anastomosis is achieved by pulling the abdominal aorta of the donor graft over the everted common carotid artery of the recipient animal. Venous drainage of the graft is achieved by pulling the portal vein of the graft over the everted external jugular vein of the recipient. This manuscript provides details and crucial steps of the organ recovery and organ implantation procedures, which will allow researchers with microsurgical skills to perform the transplantation successfully in their laboratories.

Among abdominal solid organ transplantation, pancreatic grafts are prone to develop severe ischemia reperfusion injury-associated graft damage, leading eventually to early graft loss. This protocol describes a model of murine pancreas transplantation using a non-suture cuff technique, ideally suited for analyzing these early, deleterious damages.