325:. Moreover, it is important to consider the thermal specialisation of skeletal muscles, with core muscles more susceptible to changes in mechanical performance with small temperature changes than peripheral muscles. In ectotherms (reptiles, amphibians, fish, and invertebrates), the muscle tissue can survive outside of the organism for hours or even days, depending on the temperature and organism. Many experiments are conducted at or near 0°C to prolong the usable life of the muscle. Additionally, in fish and amphibians, it is possible to separate out a single muscle fiber while keeping it intact, but in other species, this is usually not possible.
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to ensure the animal is completely dead without the possibility of recovery, which includes cessation of blood flow via the removal of the heart from the circulatory system and/or complete destruction of the brain and spinal column. Following this, common measures of animal morphology are usually rapidly obtained, such as animal length, body mass, and other biomechanical markers that may be of importance. The animal is then prepared for harvesting of the target muscle. In isolated muscles, these tend to be muscles of the hind limbs, such as the
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with the appropriate concentration of oxygen and carbon dioxide for the tissue that is being prepared. Typically, non-mammalian tissues are prepared in a gaseous solution bubbled through with 98% oxygen, 2% carbon dioxide whilst mammalian tissues in a solution bubbled through with 95% oxygen, 5% carbon dioxide. A microscope with an appropriate magnification level is required due to the dexterity required for isolation of muscles. An external, fibre optic light source is also beneficial to provide sufficient light without the emission of heat.
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rapid cell death once muscle tissue is removed from the organism. Mammalian skeletal muscles are commonly tested at ~25°C to prolong the test protocol for as long as reasonably possible. A test temperature of ~37°C can also been used during testing of whole isolated mammalian skeletal muscles to better replicate the temperature found in
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with oxygen bubbling through in order to keep the tissue alive and metabolically active. The solution is heated, usually via an external heater/cooler water bath, to an appropriate test temperature for the muscle that is being tested. Muscles are stimulated to contract by applying electric current to
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muscle testing typically requires a dual-mode servomotor, which can both control and detect changes in force and length. Should a dual-mode system be unavailable, then an independent force transducer and motor arm can be used. One end of the sample tissue is anchored in place, via a needle if sutured
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function and fibre type distribution. Following ethical approval, and if necessary, government approval, the animal is humanely euthanised. Humane methods differ by country, with the most appropriate based on ethical approval and researcher skill level. A number of further criteria should be followed
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muscle testing is almost never used in humans, with the exception of small sections of muscle removed via biopsy or while undergoing surgery for other ailments. Testing is generally more difficult in mammals and birds because of the high temperature and oxygen requirements of the muscle, leading to
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to ensure metabolic conditions are slowed down, hence the need for chilled dissecting medium, and to prevent the tissue from dying due to lack of substrates within the medium, hence the requirement for the solutions to be changed frequently. The dissecting solution should be continually oxygenated
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There is no correct approach for the preparation of muscles for testing, as long as the muscle is not damaged during preparation, the muscle-tendon unit is intact and there is something that can be used to anchor the muscle within the testing rig. Pieces of bone can be left at the proximal and/or
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tests are performed with the muscle still attached to the organism. This ensures the muscle is kept at the right temperature and amply supplied with nutrients and oxygen by the blood, but the procedure is more difficult and some tests may not be possible.
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distal end of skeletal muscles to allow for anchoring. In addition, silk sutures or aluminium T-foil clips can be used to wrap around the tendon of the muscle to provide both support at the tendon and to be used for anchoring in the mechanics rig.
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or even a half-sarcomere. Muscle fibers may be intact, or may be "skinned", a process which removes the cell membrane, sarcoplasmic reticulum, and cytoplasm, allowing greater access to the contractile components of the sarcomere.
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either the nerve which innervates the muscle or via platinum electrodes placed in the circulating solution to evoke a response of the entire muscle. The servomotor detects changes in force and/or length due to
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testing allows for exact stimulation of the muscle, providing precise data on innate tissue behavior. Isolated muscle testing limits other factors on the environment around the tissue such as substrates.
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Several properties are commonly tested, and a given experiment will often use a subset of these properties, including twitch times, tetanic force, force-length relationship, force velocity relationship,
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recruitment. The servomotor can be programmed to maintain a given force while allowing the muscle to change length, vice versa, or the muscle may be subject to more complex testing, such as in
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muscle testing has provided the bulk of scientific knowledge of muscle structure and physiology, and how both relate to organismal performance. Stem cell research relies on
527:"Regional thermal specialisation in a mammal: temperature affects power output of core muscle more than that of peripheral muscle in adult mice (Mus musculus)"
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muscle testing to establish sole muscle cell function and its individual behavior apart from muscle cells in the presence of nonmuscle cells seen in
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For the successful isolation of skeletal muscles, specific conditions are required. The tissue should be isolated in frequently changed, chilled
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or crocodile clip if prepared with aluminium T-foil clips, while the other end is attached to the servomotor. The entire muscle is bathed in
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tissue after removing it from an organism, which allows more extensive and precise quantification of its properties than
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for jumping); or a specific animal strain, to answer a research question—a specific muscle is identified based on its
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256:(provided they share a common insertion or origin, as in the human quadriceps), a single muscle, a "bundle" of
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isolated muscle testing is a beneficial procedure based on its ideal accuracy, precision, and reproducibly.
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allows individual data of muscle cell function without the presence of signaling nonmuscle cells nearby.
577:"Skeletal muscle explants: ex-vivo models to study cellular behavior in a complex tissue environment"
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626:"3D in vitro models of skeletal muscle: myopshere, myobundle and bioprinted muscle construct"
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Once an appropriate animal has been selected—whether for a specific locomotor function (i.e.
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or iliotibialis of amphibians. Other muscles that have been examined
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is often used to accurately determine fiber length during the test.
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Dessauge F, Schleder C, Perruchot MH, Rouger K (May 2021).
488:"Advantages of Testing Muscle Mechanics in-situ or in-vivo"
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429:"Isolation and characterization of human muscle cells"
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is a method used to characterize properties of living
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Testing of living muscle tissue outside of an organism
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350:Disadvantages of isolated muscle testing
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486:aursc20dev (2012-11-08).
58:"In vitro muscle testing"
333:Isolating muscle tissue
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218:Krebs-Henseleit solution
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400:Physiology
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227:motor unit
69:newspapers
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377:July 2019
262:sarcomere
204:Equipment
174:plantaris
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506:cite web
389:See also
344:In vitro
339:In vitro
335:in vitro
318:In vitro
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140:In vitro
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473:6946499
441:Bibcode
323:in vivo
313:Species
306:in vivo
302:in situ
298:in vivo
287:In situ
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530:(PDF)
245:Scale
161:frogs
90:JSTOR
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550:PMID
512:link
469:PMID
296:and
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