This work was supported by the National Health and Medical Research Council [570930]. Imaging was carried out at the Bosch Institute Advanced Microscopy Facility. Former members of the lab, whose work is cited, are thanked for their contributions to developing these methods.

<ol><li>Schmidt, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neuregulin%2FErbB+regulate+neuromuscular+junction+development+by+phosphorylation+of+%CE%B1-dystrobrevin%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Neuregulin/ErbB regulate neuromuscular junction development by phosphorylation of α-dystrobrevin.</a> J Cell Biol. 195, 1171-1184 (2011).</li>
<li>Amenta, A. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Biglycan+is+an+extracellular+MuSK+binding+protein+important+for+synapse+stability%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Biglycan is an extracellular MuSK binding protein important for synapse stability.</a> J Neurosci. 32, 2324-2334 (2012).</li>
<li>Samuel, M. A., Valdez, G., Tapia, J. C., Lichtman, J. W., Sanes, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Agrin+and+Synaptic+Laminin+Are+Required+to+Maintain+Adult+Neuromuscular+Junctions%5BTitle%5D)+AND+%22PLOS+ONE%22%5BJournal%5D)">Agrin and Synaptic Laminin Are Required to Maintain Adult Neuromuscular Junctions.</a> PLOS ONE. 7, e46663(2012).</li>
<li>Valdez, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Attenuation+of+age-related+changes+in+mouse+neuromuscular+synapses+by+caloric+restriction+and+exercise%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+%28USA%29%22%5BJournal%5D)">Attenuation of age-related changes in mouse neuromuscular synapses by caloric restriction and exercise.</a> Proc Natl Acad Sci (USA). 107, 14863-14868 (2010).</li>
<li>Yampolsky, P., Pacifici, P. G., Witzemann, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differential+muscle-driven+synaptic+remodeling+in+the+neuromuscular+junction+after+denervation%29%5BTitle%5D)+AND+%22Eur+J+Neurosci%22%5BJournal%5D)">Differential muscle-driven synaptic remodeling in the neuromuscular junction after denervation).</a> Eur J Neurosci. 31, 646-658 (2010).</li>
<li>Li, Y., Lee, Y., Thompson, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Changes+in+Aging+Mouse+Neuromuscular+Junctions+Are+Explained+by+Degeneration+and+Regeneration+of+Muscle+Fiber+Segments+at+the+Synapse%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Changes in Aging Mouse Neuromuscular Junctions Are Explained by Degeneration and Regeneration of Muscle Fiber Segments at the Synapse.</a> J Neurosci. 31, 14910-14919 (2011).</li>
<li>Zhu, H., Bhattacharyya, B. J., Lin, H., Gomez, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Skeletal+muscle+IP3R1+receptors+amplify+physiological+and+pathological+synaptic+calcium+signals%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Skeletal muscle IP3R1 receptors amplify physiological and pathological synaptic calcium signals.</a> J Neurosci. 31, 15269-15283 (2011).</li>
<li>Valdez, G., Tapia, J. C., Lichtman, J. W., Fox, M. A., Sanes, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Shared+resistance+to+aging+and+ALS+in+neuromuscular+junctions+of+specific+muscles%5BTitle%5D)+AND+%22PLoS+ONE%22%5BJournal%5D)">Shared resistance to aging and ALS in neuromuscular junctions of specific muscles.</a> PLoS ONE. 7, e34640(2012).</li>
<li>Perez-Garcia, M. J., Burden, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Increasing+MuSK+Activity+Delays+Denervation+and+Improves+Motor+Function+in+ALS+Mice%5BTitle%5D)+AND+%22Cell+reports%22%5BJournal%5D)">Increasing MuSK Activity Delays Denervation and Improves Motor Function in ALS Mice.</a> Cell reports. 2, 1-6 (2012).</li>
<li>Klooster, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle-specific+kinase+myasthenia+gravis+IgG4+autoantibodies+cause+severe+neuromuscular+junction+dysfunction+in+mice%5BTitle%5D)+AND+%22Brain%22%5BJournal%5D)">Muscle-specific kinase myasthenia gravis IgG4 autoantibodies cause severe neuromuscular junction dysfunction in mice.</a> Brain. 135, 1081-1101 (2012).</li>
<li>Pratt, S. J., Shah, S. B., Ward, C. W., Inacio, M. P., Stains, J. P., Lovering, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+in+vivo+injury+on+the+neuromuscular+junction+in+healthy+and+dystrophic+muscles%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Effects of in vivo injury on the neuromuscular junction in healthy and dystrophic muscles.</a> J Physiol. 591, 559-570 (2013).</li>
<li>Landis, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+call+for+transparent+reporting+to+optimize+the+predictive+value+of+preclinical+research%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">A call for transparent reporting to optimize the predictive value of preclinical research.</a> Nature. 490, 187-191 (2012).</li>
<li>Gervásio, O. L., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Increased+ratio+of+rapsyn+to+ACh+receptor+stabilizes+postsynaptic+receptors+at+the+mouse+neuromuscular+synapse%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Increased ratio of rapsyn to ACh receptor stabilizes postsynaptic receptors at the mouse neuromuscular synapse.</a> J Physiol. 562, 673-685 (2005).</li>
<li>Gervásio, O. L., Armson, P. F., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Developmental+increase+in+the+amount+of+rapsyn+per+acetylcholine+receptor+promotes+postsynaptic+receptor+packing+and+stability%5BTitle%5D)+AND+%22Dev+Biol%22%5BJournal%5D)">Developmental increase in the amount of rapsyn per acetylcholine receptor promotes postsynaptic receptor packing and stability.</a> Dev Biol. 305, 262-275 (2007).</li>
<li>Brockhausen, J., Cole, R. N., Gervásio, O. L., Ngo, S. T., Noakes, P. G., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Neural+agrin+increases+postsynaptic+ACh+receptor+packing+by+elevating+rapsyn+protein+at+the+mouse+neuromuscular+synapse%5BTitle%5D)+AND+%22Dev+Neurobiol%22%5BJournal%5D)">Neural agrin increases postsynaptic ACh receptor packing by elevating rapsyn protein at the mouse neuromuscular synapse.</a> Dev Neurobiol. 68, 1153-1169 (2008).</li>
<li>Cole, R. N., Reddel, S. W., Gervásio, O. L., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Anti-MuSK+patient+antibodies+disrupt+the+mouse+neuromuscular+junction%5BTitle%5D)+AND+%22Ann+Neurol%22%5BJournal%5D)">Anti-MuSK patient antibodies disrupt the mouse neuromuscular junction.</a> Ann Neurol. 63, 782-789 (2008).</li>
<li>Morsch, M., Reddel, S. W., Ghazanfari, N., Toyka, K. V., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+Specific+Kinase+autoantibodies+cause+synaptic+failure+through+progressive+wastage+of+postsynaptic+acetylcholine+receptors%5BTitle%5D)+AND+%22Exp+Neurol%22%5BJournal%5D)">Muscle Specific Kinase autoantibodies cause synaptic failure through progressive wastage of postsynaptic acetylcholine receptors.</a> Exp Neurol. 237, 237-286 (2012).</li>
<li>Cole, R. N., Ghazanfari, N., Ngo, S. T., Gervasio, O. L., Reddel, S. W., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Patient+autoantibodies+deplete+postsynaptic+Muscle+Specific+Kinase+leading+to+disassembly+of+the+ACh+receptor+scaffold+and+myasthenia+gravis+in+mice%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Patient autoantibodies deplete postsynaptic Muscle Specific Kinase leading to disassembly of the ACh receptor scaffold and myasthenia gravis in mice.</a> J Physiol. 588, 3217-3229 (2010).</li>
<li>Viegas, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Passive+and+active+immunization+models+of+MuSK-Ab+positive+myasthenia%3A+Electrophysiological+evidence+for+pre+and+postsynaptic+defects%5BTitle%5D)+AND+%22Exp+Neurol%22%5BJournal%5D)">Passive and active immunization models of MuSK-Ab positive myasthenia: Electrophysiological evidence for pre and postsynaptic defects.</a> Exp Neurol. 234, 506-512 (2012).</li>
<li>Morsch, M., Reddel, S. W., Ghazanfari, N., Toyka, K. V., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pyridostigmine+but+not+3%2C4-diaminopyridine+exacerbates+ACh+receptor+loss+and+myasthenia+induced+in+mice+by+Muscle+Specific+Kinase+autoantibody%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Pyridostigmine but not 3,4-diaminopyridine exacerbates ACh receptor loss and myasthenia induced in mice by Muscle Specific Kinase autoantibody.</a> J Physiol. 591, 2747-2762 (2013).</li>
<li>Ghazanfari, N., Morsch, M., Reddel, S. W., Liang, S. X., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+Specific+Kinase+autoantibodies+suppress+the+MuSK+pathway+and+ACh+receptor+retention+at+the+mouse+neuromuscular+junction%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Muscle Specific Kinase autoantibodies suppress the MuSK pathway and ACh receptor retention at the mouse neuromuscular junction.</a> J Physiol. 592, 2881-2897 (2014).</li>
<li>Cheng, A., Morsch, M., Murata, Y., Ghazanfari, N., Reddel, S. W., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Sequence+of+age-associated+changes+to+the+mouse+neuromuscular+junction+and+the+protective+effects+of+voluntary+exercise%5BTitle%5D)+AND+%22PLoS+One%22%5BJournal%5D)">Sequence of age-associated changes to the mouse neuromuscular junction and the protective effects of voluntary exercise.</a> PLoS One. 8, e67970(2013).</li>
<li>Schaefer, A. M., Sanes, J. R., Lichtman, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+compensatory+subpopulation+of+motor+neurons+in+a+mouse+model+of+amyotrophic+lateral+sclerosis%5BTitle%5D)+AND+%22J+Comp+Neurol%22%5BJournal%5D)">A compensatory subpopulation of motor neurons in a mouse model of amyotrophic lateral sclerosis.</a> J Comp Neurol. 490, 209-219 (2005).</li>
<li>Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M., Altman, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improving+bioscience+research+reporting%3A+the+ARRIVE+guidelines+for+reporting+animal+research%5BTitle%5D)+AND+%22PLos+Biol%22%5BJournal%5D)">Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research.</a> PLos Biol. 8, e1000412(2010).</li>
<li>Shimizu, S. Routes of Administration. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aShimizu%2C+S+%22The+Laboratory+Mouse%22">The Laboratory Mouse</a>. Hedrich, H. J., Bullock, G. , Elsevier. (2004).</li>
<li>Chiasson, R. B. Laboratory anatomy of the white rat. , Brown. Dubuque, Iowa. (1988).</li>
<li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Whole+Animal+Perfusion+Fixation+for+Rodents%5BTitle%5D)+AND+%22J.+Vis.+Exp%22%5BJournal%5D)">Whole Animal Perfusion Fixation for Rodents.</a> J. Vis. Exp. (65), e3564(2012).</li>
<li>Mitra, A. K., Stroud McCarthy, M. P., M, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Three-dimensional+structure+of+the+nicotinic+acetylcholine+receptor+and+location+of+the+major+associated+43-kD+cytoskeletal+protein%2C+determined+at+22A+by+low+dose+electron+microscopy+and+x-ray+diffraction+to+12.5A%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Three-dimensional structure of the nicotinic acetylcholine receptor and location of the major associated 43-kD cytoskeletal protein, determined at 22A by low dose electron microscopy and x-ray diffraction to 12.5A.</a> J Cell Biol. 109, 755-774 (1989).</li>
<li>Paas, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Electron+microscopic+evidence+for+nucleation+and+growth+of+3D+acetylcholine+receptor+microcrystals+in+structured+lipid-detergent+matrices%5BTitle%5D)+AND+%22Proc.+Natl+Acad.+Sci.+%28USA%29%22%5BJournal%5D)">Electron microscopic evidence for nucleation and growth of 3D acetylcholine receptor microcrystals in structured lipid-detergent matrices.</a> Proc. Natl Acad. Sci. (USA). 100, 11309-11314 (2003).</li>
<li>Samson, A. O., Scherf, T., Eisenstein, M., Chill, J. H., Anglister, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+mechanism+for+acetylhcoline+receptor+inhibition+by+%CE%B1-neurotoxins+and+species-specific+resistance+to+%CE%B1-bungarotoxin+revealed+by+NMR%29%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">The mechanism for acetylhcoline receptor inhibition by α-neurotoxins and species-specific resistance to α-bungarotoxin revealed by NMR).</a> Neuron. 35, 319-332 (2002).</li>
<li>Ghazanfari, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Muscle+Specific+Kinase%3A+Organiser+of+synaptic+membrane+domains%5BTitle%5D)+AND+%22Int+J+Biochem+Cell+Biol%22%5BJournal%5D)">Muscle Specific Kinase: Organiser of synaptic membrane domains.</a> Int J Biochem Cell Biol. 43, 295-298 (2011).</li>
<li>Ghazanfari, N., Morsch, M., Tse, N., Reddel, S. W., Phillips, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+the+%CE%B22-adrenoceptor+agonist%2C+albuterol%2C+in+a+mouse+model+of+anti-MuSK+myasthenia+gravis%5BTitle%5D)+AND+%22PLoS+ONE%22%5BJournal%5D)">Effects of the β2-adrenoceptor agonist, albuterol, in a mouse model of anti-MuSK myasthenia gravis.</a> PLoS ONE. 9, e87840(2014).</li>
<li>Prakash, Y. S., Miller, S. M., Huang, M., Sieck, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Morphology+of+diaphragm+neuromuscular+junctions+on+different+fibre+types%5BTitle%5D)+AND+%22J+Neurocytol%22%5BJournal%5D)">Morphology of diaphragm neuromuscular junctions on different fibre types.</a> J Neurocytol. 25, 88-100 (1996).</li>
<li>Salpeter, M. M., Harris, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Distribution+and+turnover+rate+of+acetylcholine+receptors+throughout+the+junction+folds+at+a+vertebrate+neuromuscular+junction%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Distribution and turnover rate of acetylcholine receptors throughout the junction folds at a vertebrate neuromuscular junction.</a> J Cell Biol. 96, 1781-1785 (1983).</li>
<li>Soper, S. A., Nutter, H. L., Keller, R. A., Davis, L. M., Shera, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+photophysical+constants+of+several+fluorescent+dyes+pertaining+to+ultrasensitive+fluorescence+spectroscopy%5BTitle%5D)+AND+%22Photochem+Photobiol%22%5BJournal%5D)">The photophysical constants of several fluorescent dyes pertaining to ultrasensitive fluorescence spectroscopy.</a> Photochem Photobiol. 57, 972-977 (1993).</li>
<li>Panchuk-Voloshina, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Alexa+dyes%2C+a+series+of+new+fluorescent+dyes+that+yield+exceptionally+bright%2C+photostable+conjugates%5BTitle%5D)+AND+%22J+Histochem+Cytochem%22%5BJournal%5D)">Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates.</a> J Histochem Cytochem. 47, 1179-1188 (1999).</li>
</ol>

The authors declare they have no competing financial interests.

여기에 설명 된 프로토콜은 신뢰성있게 측정하고 정상 노화와 질병 상태를 포함한 조건의 범위에 걸쳐 NMJ의 특성의 변화를 정량화 할 수있게했다. EN에 대해 기재된 방법 NMJ 이미지 연구자 사전 및 시냅스 전문 분야의 오버랩 영역과 연접 / 배향의 면적과 비교 할 수 직면한다. 사전 및 시냅스 단백질 횡단 섹션을 사용하여 광 제 2 프로토콜의 상대 강도를 비교하기 위해 바람직하다. 세 번째 프로?...

신경 근육 접합 (NMJ)는 신경계와 골격 근육 사이의 통신을 매개하는 중요한 릴레이 시냅스이다. 그것은 모든 자발적인 운동이 필요합니다. 형광 현미경 긴 NMJ 1-3 마우스 유전자의 효과를 연구하기 위해 또는 설치류 4-11 NMJs시 연령,식이 요법, 운동 및 질환의 효과를 비교하기 위해 사용되었다. 이러한 연구는 NMJ의 생리학 및 병리학 소개 훨씬 가르친하지만 다양한 매개 변수 (예, ACHR 영역 종판 영역, 경계 길이, 분열 인덱스)가 종종 어렵게 이들 연구의 결과를 비교할 수 있도록보고. 전임상 연구는 특히 질병 (12)의 설치류 모델 연구에서, 재현성을 입증 할 수 있으려면 증가 기대가있다. 여기에 설명 된 프로토콜은, 생리 학적 및 병태 생리 학적 발달 CH 조사 일련의 연구를 통해 정제시켰다NMJ에 ANGES. 이러한 연구는 마우스 모터 종판에서 시냅스 전문화 영역과 시냅스 전문화 13-15 시냅스 내의 단백질의 패킹의 상대 밀도의 측정을 필요로한다. 이 방법의 유용성은 중증 근무력증 안티 사향 마우스 모델에서 최근 연구에 의해 예시된다. 안티 사향 양성 중증에서의 IgG 매일 ​​주사는 성인 마우스로 환자들을 2 주 (16) 내에서 약이 될 인한 근무력증. 근육 (신경 터미널에서) synaptophysin에 대한 이중 표지 된 섹션과 시냅스 AChRs의 공 초점 최대 프로젝션 이미지가 주 변화로 ACHR 염색 분야의 진보적 인 감소를 한 것으로 밝혀졌습니다. 감소의 중요한 비율은 시냅스 전위, 시냅스 전달 및 근육 약화 (17, 18)의 실패의 진폭에 비교 하락을 설명하기에 충분했다. 질적으로 비슷한 연구 결과는 다른 연구 그룹에 의해보고되었다10,19. NMJ 동일한 측정 방법은 사람이 마우스 모델 (20, 21)에 근무력증 안티 사향 중증의 치료 세 약물의 영향을 평가하기 위해 사용되어왔다. 정주의 노화는 신경 근육 연결의 손실을 초래할 수 있습니다. 마우스는 나이로 진행하면서 여기에 설명 된 프로토콜은 모터 종판에서 신경 터미널 시납의 영역에서 연령 관련 감소를 계시했다. 같은 방법은 자발적인 운동이 크게 다른 그룹 4로 이전의 연구와 일치하는 시냅스 전 신경 말단 영역 (22)의 감소를 방지 할 수있는 것으로 나타났습니다. 신경 근육 연결의 손실은 근 위축성 측삭 경화증 9,23의 SOD1G93A 마우스 모델에서 발생합니다. 상기 언급 된 연구들은 건강 상태의 다양한 NMJ에서 전후 시냅스 전문 하나의 영역에서 감소를 초래할 수 있음을 입증한다. 이것은 손상된 시냅스 재미가 발생할 수 있습니다ction의는 신경 근육 연결의 완전한 손실을 예고하거나. 세 가지 프로토콜은 지역과 시냅스 전문의 밀도의 정량을 허용하는 설명되어 있습니다. 제 1 프로토콜의 목적은 형광 현미경을 이용하여, 전후 및 포유 동물의 시냅스 전문 NMJs 그들의 배향의 영역의 실제적이고 재현 가능한 측정 값을 제공하는 것이다. 이차원 최대 투영 공 초점 이미지 및 NIH ImageJ에 함께 이미지 분석 시납 염색 (시냅스 소포), 시냅스 AChRs 시냅스 오버랩 영역의 면적의 변화를 감지하는 데 사용된다. 시냅스 전문 영역을 식별하는 데 사용되는 시각 정보를 최대화하기 위해 공 초점 촬상 파라미터 (이득 및 오프셋 레벨)이 각각 NMJ 최적화되어있다. 신경 근육 장애는 시냅스 ACHR 및 / 또는 기타 시냅스 단백질의 밀도의 변화로 발생할 수 있습니다. 제 2 프로토콜은 시냅스 단백질 등의 상대 밀도의 변화를 검출하도록 적용될 수있다사향, rapsyn, dystroglycan, 인산화의 Src 키나제 인산화 ACHR 18,21있다. 중증 근무력증에서 시냅스 막 내 ACHR의 감소 밀도는 시냅스 실패와 근육 약화의 직접적인 원인이다. 세 번째 프로토콜은 시냅스 막 (14, 15) 내에서 인접 AChRs의 근접의 변화를 평가하기 위해 형광 공명 에너지 전달 (FRET) 방법을 설명합니다. 이 방법은 형광-α-bungarotoxin (BGT)으로 표시 이웃 AChRs 사이의 에너지 전달을 검출한다. 형광 기증자 및 수용체 프로브는 10nm 이하 떨어져있는 경우 FRET가 발생합니다. 이것은 직접 시냅스 전위의 진폭에 관련 될 수있다 ACHR 포장의 압박감에서 (초 현미경) 변경을 표시 할 수 있습니다. 지난 10 년 동안 정제 된이 세 가지 프로토콜은 일관되고 재현 가능한 방식으로 NMJ 무결성의 보완을 제공합니다. 표준화 된 프로토콜의 사용ND 파라미터 NMJ 포유류 유전자, 환경에 따라 중재 효과 비교를 용이하게한다.

<table><tbody><tr><td>Scanning confocal microscope</td><td>Leica</td><td>DM 2000 with&amp;nbsp; TCS SP2 system</td><td>Most scanning confocal microscopes should be suitable.&amp;nbsp;</td></tr><tr><td></td><td>Zeiss</td><td>LSM 510 Meta&amp;nbsp;</td><td></td></tr><tr><td></td><td>Leica</td><td>SPE-II</td><td></td></tr><tr><td>Alexa555-a-bungarotoxin (red-BGT)</td><td>Life technologies</td><td>B35451</td><td>Used for labelling AChRs</td></tr><tr><td>Alexa647-&amp;alpha;-bungarotoxin (far-red-BGT)</td><td>Life technologies</td><td>B35450</td><td>Far red fluorescence: barely visible through the eyepiece&amp;nbsp;</td></tr><tr><td>rabbit anti-synaptophysin</td><td>Life technologies</td><td>18-0130</td><td>Different batches of primary antibody differ in effective working dilution</td></tr><tr><td>FITC-anti-rapsyn mab1234</td><td>Milipore</td><td>FCMAB134F</td><td>Monoclonal antibody conjugated to FITC</td></tr><tr><td>FITC-donkey anti-rabbit IgG</td><td>Jackson</td><td>711-095-152</td><td>Polyclonal secondary antibodies can vary in quality according to source and batch</td></tr><tr><td>Optimal Cutting Temperature compound (O.T.C.)</td><td>ProSciTech</td><td>IA018</td><td>Cryostat embedding matrix for freezing&amp;nbsp; muscles</td></tr><tr><td>DABCO</td><td>Sigma</td><td>10981</td><td>Mounting medium that slows photobleaching of fluorophores</td></tr></tbody></table>

the neuromuscular junction: measuring synapse size, fragmentation and changes in synaptic protein density using confocal fluorescence microscopy

신경 근육 접합 (NMJ)은 포유 동물의 운동 신경이 자발적 근육 수축을 제어되는 큰, 콜린성 릴레이 시냅스이다. NMJ에서 구조 변경 약화, 위축과 근육 섬유의 사망을 초래, 신경 전달 오류가 발생할 수 있습니다. 많은 연구는 유전자 변형 또는 질환 마우스 NMJ의 구조를 변경할 수있는 방법을 연구 하였다. 불행히도, 직접들은 종종 다른 파라미터 분석 방법을 사용하기 때문에 이들 연구에서 조사 결과를 비교하기 어려울 수있다. 세 개의 프로토콜이 설명되어 있습니다. 제는 아세틸 콜린 수용체 (ACHR)의​​ 면적을 측정하는 최대 강도 투영 공 촛점 이미지를 사용하여 단부 판과 상부 시냅스 전 신경 말단의 시냅스 소포 염색이 풍부한 영역에서 시냅스 막 도메인. 두 번째 프로토콜은 시냅스 막 시냅스 단백질에 대한 면역 염색의 상대적 강도를 비교합니다. 세 번째 홍보otocol은 엔드 플레이트에서 시냅스 AChRs의 포장의 변화를 감지하는 형광 공명 에너지 전달 (FRET)를 사용합니다. 프로토콜이 개발 및 일련의 연구를 통해 정제되었다. 품질과 결과의 일관성에 영향을 미치는 요인에 대해 설명하고 규범 적 데이터는 건강한 젊은 성인 마우스의 NMJs 제공됩니다.

NMJ에서 시냅스 지역 측정 영역의 모든 추정치는 시냅스 전문의 범위를 정의하는 경계의 도면에 의존. 건강한 젊은 성인 근육에 NMJ 이미지 ACHR과 synaptophysin에 염색 (그림 2A와 B) 모두 잘 정의 된 경계를 표시해야합니다. ACHR 및 시납 모두 형광 강도는 모터 엔드 플레이트 (도 5a &#39;및 B&#39;)의-주위염 시냅스와 시냅스 사이의 경...

Watch this Scientific Journal Video about The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy at JoVE.com

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

신경 근육 접합 : 측정 시냅스 크기, 공 초점 형광 현미경을 사용하여 시냅스 단백질 농도의 분열 및 변경

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

the-neuromuscular-junction-measuring-synapse-size-fragmentation

Research

JoVE Journal

Neuroscience

21.6K 조회수.  University of Sydney. The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

비디오: The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number

One of the most important goals in neuroscience is to understand the molecular cues that instruct early stages of synapse formation. As such it has become imperative to develop objective approaches to quantify changes in synaptic connectivity. Starting from sample fixation, this protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry. Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization. We define synapses as points of colocalization between the signals generated by these markers. The number of these colocalizations is quantified using a plug in Puncta Analyzer (written by Bary Wark, available upon request, c.eroglu@cellbio.duke.edu) under the ImageJ analysis software platform. The synapse assay described in this protocol can be applied to any neural tissue or culture preparation for which you have selective pre- and postsynaptic markers. This synapse assay is a valuable tool that can be widely utilized in the study of synaptic development.

This protocol details how to quantify synapse number both in dissociated neuronal culture and in brain sections using immunocytochemistry. Using compartment-specific antibodies, we label presynaptic terminals as well as sites of postsynaptic specialization. We define synapses as points of colocalization between the signals generated by these markers.

Quantifying 시냅스 : 시냅스 번호를 수치 Immunocytochemistry 기반 어세이

One of the most important goals in neuroscience is to understand the molecular cues that instruct early stages of synapse formation. As such it has become ...

연구

신경과학

Analysis of Dendritic Spine Morphology in Cultured CNS Neurons

Dendritic spines are the sites of the majority of excitatory connections within the brain, and form the post-synaptic 
compartment of synapses. These structures are rich in actin and have been shown to be highly dynamic. In response to classical Hebbian plasticity 
as well as neuromodulatory signals, dendritic spines can change shape and number, which is thought to be critical for the refinement of neural 
circuits and the processing and storage of information within the brain. Within dendritic spines, a complex network of proteins link extracellular 
signals with the actin cyctoskeleton allowing for control of dendritic spine morphology and number. Neuropathological studies have demonstrated that 
a number of disease states, ranging from schizophrenia to autism spectrum disorders, display abnormal dendritic spine morphology or numbers. 
Moreover, recent genetic studies have identified mutations in numerous genes that encode synaptic proteins, leading to suggestions that these 
proteins may contribute to aberrant spine plasticity that, in part, underlie the pathophysiology of these disorders. In order to study the potential 
role of these proteins in controlling dendritic spine morphologies/number, the use of cultured cortical neurons offers several advantages. Firstly, 
this system allows for high-resolution imaging of dendritic spines in fixed cells as well as time-lapse imaging of live cells. Secondly, this in 
vitro system allows for easy manipulation of protein function by expression of mutant proteins, knockdown by shRNA constructs, or pharmacological 
treatments. These techniques allow researchers to begin to dissect the role of disease-associated proteins and to predict how mutations of these 
proteins may function in vivo.

Numerous recent studies have identified mutations in synaptic proteins associated with brain pathologies. Primary cultured cortical neurons offer great flexibility in examining the effects of these disease-associated proteins on dendritic spine morphology and motility.

교양 CNS의 뉴런의 돌기 스파인 형태론 분석

Dendritic spines are the sites of the majority of excitatory connections within the brain, and form the post-synaptic 
compartment of synapses. These ...

Automated Quantification of Synaptic Fluorescence in C. elegans

Synapse strength refers to the amplitude of postsynaptic responses to presynaptic neurotransmitter release events, and has a major impact on overall neural circuit function. Synapse strength critically depends on the abundance of neurotransmitter receptors clustered at synaptic sites on the postsynaptic membrane. Receptor levels are established developmentally, and can be altered by receptor trafficking between surface-localized, subsynaptic, and intracellular pools, representing important mechanisms of synaptic plasticity and neuromodulation. Rigorous methods to quantify synaptically-localized neurotransmitter receptor abundance are essential to study synaptic development and plasticity. Fluorescence microscopy is an optimal approach because it preserves spatial information, distinguishing synaptic from non-synaptic pools, and discriminating among receptor populations localized to different types of synapses. The genetic model organism Caenorhabditis elegans is particularly well suited for these studies due to the small size and relative simplicity of its nervous system, its transparency, and the availability of powerful genetic techniques, allowing examination of native synapses in intact animals.
Here we present a method for quantifying fluorescently-labeled synaptic neurotransmitter receptors in C. elegans. Its key feature is the automated identification and analysis of individual synapses in three dimensions in multi-plane confocal microscope output files, tabulating position, volume, fluorescence intensity, and total fluorescence for each synapse. This approach has two principal advantages over manual analysis of z-plane projections of confocal data. First, because every plane of the confocal data set is included, no data are lost through z-plane projection, typically based on pixel intensity averages or maxima. Second, identification of synapses is automated, but can be inspected by the experimenter as the data analysis proceeds, allowing fast and accurate extraction of data from large numbers of synapses. Hundreds to thousands of synapses per sample can easily be obtained, producing large data sets to maximize statistical power. Considerations for preparing C. elegans for analysis, and performing confocal imaging to minimize variability between animals within treatment groups are also discussed. Although developed to analyze C. elegans postsynaptic receptors, this method is generally useful for any type of synaptically-localized protein, or indeed, any fluorescence signal that is localized to discrete clusters, puncta, or organelles.
The procedure is performed in three steps: 1) preparation of samples, 2) confocal imaging, and 3) image analysis. Steps 1 and 2 are specific to C. elegans, while step 3 is generally applicable to any punctate fluorescence signal in confocal micrographs.

Automated Quantification of Synaptic Fluorescence in C. elegans

The abundance of neurotransmitter receptors clustered at synapses strongly influences synaptic strength. This method quantifies fluorescently-labeled neurotransmitter receptors in three dimensions with single-synapse resolution in C. elegans, allowing hundreds of synapses to be rapidly characterized within a single sample without distortions introduced by z-plane projection.

에서 시냅틱 형광의 자동 부량<em&gt; C. elegans</em

Synapse strength refers to the amplitude of postsynaptic responses to presynaptic neurotransmitter release events, and has a major impact on overall ...

DetectSyn: A Rapid, Unbiased Fluorescent Method to Detect Changes in Synapse Density

Synapses are the site of communication between neurons. Neuronal circuit strength is related to synaptic density, and the breakdown of synapses is characteristic of disease states like major depressive disorder (MDD) and Alzheimer's disease. Traditional techniques to investigate synapse numbers include genetic expression of fluorescent markers (e.g., green fluorescent protein (GFP)), dyes that fill a neuron (e.g., carbocyanine dye, DiI), and immunofluorescent detection of spine markers (e.g., postsynaptic density 95 (PSD95)). A major caveat to these proxy techniques is that they only identify postsynaptic changes. Yet, a synapse is a connection between a presynaptic terminal and a postsynaptic spine. The gold standard for measuring synapse formation/elimination requires time-consuming electron microscopy or array tomography techniques. These techniques require specialized training and costly equipment. Further, only a limited number of neurons can be assessed and are used to represent changes to an entire brain region. DetectSyn is a rapid fluorescent technique that identifies changes to synapse formation or elimination due to a disease state or drug activity. DetectSyn utilizes a rapid proximity ligation assay to detect juxtaposed pre- and postsynaptic proteins and standard fluorescent microscopy, a technique readily available to most laboratories. Fluorescent detection of the resulting puncta allows for quick and unbiased analysis of experiments. DetectSyn provides more representative results than electron microscopy because larger areas can be analyzed than a limited number of fluorescent neurons. Moreover, DetectSyn works for in vitro cultured neurons and fixed tissue slices. Finally, a method is provided to analyze the data collected from this technique. Overall, DetectSyn offers a procedure for detecting relative changes in synapse density across treatments or disease states and is more accessible than traditional techniques.

DetectSyn is an unbiased, rapid fluorescent assay that measures changes in relative synapse (pre- and postsynaptic engagement) number across treatments or disease states. This technique utilizes a proximity ligation technique that can be used both in cultured neurons and fixed tissue.

DetectSyn: Synapse 밀도의 변화를 감지하는 빠르고 편향되지 않은 형광 방법

Synapses are the site of communication between neurons. Neuronal circuit strength is related to synaptic density, and the breakdown of synapses is ...

생화학

Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology

Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The Drosophila larval neuromuscular junction (NMJ), a well-established model for glutamatergic synapses, has been extensively studied for decades. Identification of mutations causing NMJ morphological defects revealed a repertoire of genes that regulate synapse development and function. Many of these were identified in large-scale studies that focused on qualitative approaches to detect morphological abnormalities of the Drosophila NMJ. A drawback of qualitative analyses is that many subtle players contributing to NMJ morphology likely remain unnoticed. Whereas quantitative analyses are required to detect the subtler morphological differences, such analyses are not yet commonly performed because they are laborious. This protocol describes in detail two image analysis algorithms "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics", available as Fiji-compatible macros, for quantitative, accurate and objective morphometric analysis of the Drosophila NMJ. This methodology is developed to analyze NMJ terminals immunolabeled with the commonly used markers Dlg-1 and Brp. Additionally, its wider application to other markers such as Hrp, Csp and Syt is presented in this protocol. The macros are able to assess nine morphological NMJ features: NMJ area, NMJ perimeter, number of boutons, NMJ length, NMJ longest branch length, number of islands, number of branches, number of branching points and number of active zones in the NMJ terminal.

Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology

Two image analysis algorithms, "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics" were created, to automatically quantify nine morphological features of the Drosophila neuromuscular junction (NMJ).

높은 처리량 및 다중 파라 메트릭 정량화를위한 ​​두 알고리즘<em&gt; 초파리</em&gt; 신경 근육 접합 형태학

Synaptic morphology is tightly related to synaptic efficacy, and in many cases morphological synapse defects ultimately lead to synaptic malfunction. The ...

Dissection of Single Skeletal Muscle Fibers for Immunofluorescent and Morphometric Analyses of Whole-Mount Neuromuscular Junctions

The neuromuscular junction (NMJ) is a specialized point of contact between the motor nerve and the skeletal muscle. This peripheral synapse exhibits high morphological and functional plasticity. In numerous nervous system disorders, NMJ is an early pathological target resulting in neurotransmission failure, weakness, atrophy, and even in muscle fiber death. Due to its relevance, the possibility to quantitatively assess certain aspects of the relationship between NMJ components can help to understand the processes associated with its assembly/disassembly. The first obstacle when working with muscles is to gain the technical expertise to quickly identify and dissect without damaging their fibers. The second challenge is to utilize high-quality detection methods to obtain NMJ images that can be used to perform quantitative analysis. This article presents a step-by-step protocol for dissecting extensor digitorum longus and soleus muscles from rats. It also explains the use of immunofluorescence to visualize pre and postsynaptic elements of whole-mount NMJs. Results obtained demonstrate that this technique can be used to establish the microscopic anatomy of the synapsis and identify subtle changes in the status of some of its components under physiological or pathological conditions.

The ability to accurately detect neuromuscular junction components is crucial in evaluating modifications in its architecture because of pathological or developmental processes. Here we present a complete description of a straightforward method to obtain high-quality images of whole-mount neuromuscular junctions that can be used to perform quantitative measurements.

면역형광및 전체산 신경근육 접합의 형태 분석에 대한 단일 골격 근육 섬유의 해부

The neuromuscular junction (NMJ) is a specialized point of contact between the motor nerve and the skeletal muscle. This peripheral synapse exhibits high ...

Real-Time Fluorescent Measurement of Synaptic Functions in Models of Amyotrophic Lateral Sclerosis

Before neuronal degeneration, the cause of motor and cognitive deficits in patients with amyotrophic lateral sclerosis (ALS) and/or frontotemporal lobe dementia (FTLD) is dysfunction of communication between neurons and motor neurons and muscle. The underlying process of synaptic transmission involves membrane depolarization-dependent synaptic vesicle fusion and the release of neurotransmitters into the synapse. This process occurs through localized calcium influx into the presynaptic terminals where synaptic vesicles reside. Here, the protocol describes fluorescence-based live-imaging methodologies that reliably report depolarization-mediated synaptic vesicle exocytosis and presynaptic terminal calcium influx dynamics in cultured neurons.
Using a styryl dye that is incorporated into synaptic vesicle membranes, the synaptic vesicle release is elucidated. On the other hand, to study calcium entry, Gcamp6m is used, a genetically encoded fluorescent reporter. We employ high potassium chloride-mediated depolarization to mimic neuronal activity. To quantify synaptic vesicle exocytosis unambiguously, we measure the loss of normalized styryl dye fluorescence as a function of time. Under similar stimulation conditions, in the case of calcium influx, Gcamp6m fluorescence increases. Normalization and quantification of this fluorescence change are performed in a similar manner to the styryl dye protocol. These methods can be multiplexed with transfection-based overexpression of fluorescently tagged mutant proteins. These protocols have been extensively used to study synaptic dysfunction in models of FUS-ALS and C9ORF72-ALS, utilizing primary rodent cortical and motor neurons. These protocols easily allow for rapid screening of compounds that may improve neuronal communication. As such, these methods are valuable not only for the study of ALS but for all areas of neurodegenerative and developmental neuroscience research.

Two related methods are described to visualize subcellular events required for synaptic transmission. These protocols enable the real-time monitoring of the dynamics of presynaptic calcium influx and synaptic vesicle membrane fusion using live-cell imaging of in vitro cultured neurons.

근위축성 측삭 경화증 모델에서 시냅스 기능의 실시간 형광 측정

Before neuronal degeneration, the cause of motor and cognitive deficits in patients with amyotrophic lateral sclerosis (ALS) and/or frontotemporal lobe ...

Characterization of Neuromuscular Junctions in Mice by Combined Confocal and Super-Resolution Microscopy

Neuromuscular junctions (NMJs) are highly specialized synapses between lower motor neurons and skeletal muscle fibers that play an essential role in the transmission of molecules from the nervous system to voluntary muscles, leading to contraction. They are affected in many human diseases, including inherited neuromuscular disorders such as Duchenne muscular dystrophy (DMD), congenital myasthenic syndromes (CMS), spinal muscular atrophy (SMA), and amyotrophic lateral sclerosis (ALS). Therefore, monitoring the morphology of neuromuscular junctions and their alterations in disease mouse models represents a valuable tool for pathological studies and preclinical assessment of therapeutic approaches. Here, methods for labeling and analyzing the three-dimensional (3D) morphology of the pre- and postsynaptic parts of motor endplates from murine teased muscle fibers are described. The procedures to prepare samples and measure NMJ volume, area, tortuosity and axon terminal morphology/occupancy by confocal imaging, and the distance between postsynaptic junctional folds and acetylcholine receptor (AChR) stripe width by super-resolution stimulated emission depletion (STED) microscopy are detailed. Alterations in these NMJ parameters are illustrated in mutant mice affected by SMA and CMS.

This protocol describes a method for morphometric analysis of neuromuscular junctions by combined confocal and STED microscopy that is used to quantify pathological changes in mouse models of SMA and ColQ-related CMS.

결합된 공초점 및 초분해능 현미경에 의한 마우스의 신경근 접합부 특성 분석

Neuromuscular junctions (NMJs) are highly specialized synapses between lower motor neurons and skeletal muscle fibers that play an essential role in the ...

Registration of Calcium Transients in Mouse Neuromuscular Junction with High Temporal Resolution using Confocal Microscopy

Estimation of the presynaptic calcium level is a key task in studying synaptic transmission since calcium entry into the presynaptic cell triggers a cascade of events leading to neurotransmitter release. Moreover, changes in presynaptic calcium levels mediate the activity of many intracellular proteins and play an important role in synaptic plasticity. Studying calcium signaling is also important for finding ways to treat neurodegenerative diseases. The neuromuscular junction is a suitable model for studying synaptic plasticity, as it has only one type of neurotransmitter. This article describes the method for loading a calcium-sensitive dye through the cut nerve bundle into the mice's motor nerve endings. This method allows the estimation of all parameters related to intracellular calcium changes, such as basal calcium level and calcium transient. Since the influx of calcium from the cell exterior into the nerve terminals and its binding/unbinding to the calcium-sensitive dye occur within the range of a few milliseconds, a speedy imaging system is required to record these events. Indeed, high-speed cameras are commonly used for the registration of fast calcium changes, but they have low image resolution parameters. The protocol presented here for recording calcium transient allows extremely good spatial-temporal resolution provided by confocal microscopy.

The protocol describes the method of loading a fluorescent calcium dye through the cut nerve into mouse motor nerve terminals. In addition, a unique method for recording fast calcium transients in the peripheral nerve endings using confocal microscopy is presented.

Estimation of the presynaptic calcium level is a key task in studying synaptic transmission since calcium entry into the presynaptic cell triggers a ...

Visualizing the Morphological Characteristics of Neuromuscular Junction in Rat Medial Gastrocnemius Muscle

The neuromuscular junction (NMJ) is a complex structure serving for the signal communication from the motor neuron to the skeletal muscle and consists of three essential histological components: the pre-synaptic motor axon terminals, post-synaptic nicotinic acetylcholine receptors (AchRs), and peri-synaptic Schwann cells (PSCs). In order to demonstrate the morphological characteristics of NMJ, the rat medial gastrocnemius muscle was selected as the target-tissue and examined by using multiple fluorescent staining with various kinds of biomarkers, including neurofilament 200 (NF200) and vesicular acetylcholine transporter (VAChT) for the motor nerve fibers and their pre-synaptic terminals, alpha-bungarotoxin (&#945;-BTX) for the post-synaptic nicotinic AchRs, and S100 for the PSCs. In this study, staining was performed in two groups: in the first group, samples were stained with NF200, VAChT, and &#945;-BTX, and in the second group, samples were stained with NF200, &#945;-BTX, and S100. It was shown that both protocols can effectively demonstrate the detailed structure of NMJ. Using the confocal microscope, morphological characteristics of the pre-synaptic terminals, post-synaptic receptors, and PSC were seen, and their Z-stacks images were reconstructed in a three-dimensional pattern to further analyze the spatial correlation among the different labeling. From the perspective of methodology, these protocols provide a valuable reference for investigating the morphological characteristics of NMJ under physiological conditions, which may also be suitable to evaluate the pathological alteration of NMJ, such as peripheral nerve injury and regeneration.

The protocol shows a method to examine spatial correlation among the pre-synaptic terminals, post-synaptic receptors, and peri-synaptic Schwann cells in the rat medial gastrocnemius muscle using fluorescent immunohistochemistry with different biomarkers, namely, neurofilament 200, vesicular acetylcholine transporter, alpha-bungarotoxin, and S100.

쥐 내측 위장근에서 신경근 접합부의 형태학적 특성 시각화

The neuromuscular junction (NMJ) is a complex structure serving for the signal communication from the motor neuron to the skeletal muscle and consists of ...

신경 근육 접합 : 측정 시냅스 크기, 공 초점 형광 현미경을 사용하여 시냅스 단백질 농도의 분열 및 변경

요약

더 많은 비디오 탐색

Quantifying 시냅스 : 시냅스 번호를 수치 Immunocytochemistry 기반 어세이

교양 CNS의 뉴런의 돌기 스파인 형태론 분석

에서 시냅틱 형광의 자동 부량

DetectSyn: Synapse 밀도의 변화를 감지하는 빠르고 편향되지 않은 형광 방법

높은 처리량 및 다중 파라 메트릭 정량화를위한 두 알고리즘

면역형광및 전체산 신경근육 접합의 형태 분석에 대한 단일 골격 근육 섬유의 해부

근위축성 측삭 경화증 모델에서 시냅스 기능의 실시간 형광 측정

결합된 공초점 및 초분해능 현미경에 의한 마우스의 신경근 접합부 특성 분석

Registration of Calcium Transients in Mouse Neuromuscular Junction with High Temporal Resolution using Confocal Microscopy

쥐 내측 위장근에서 신경근 접합부의 형태학적 특성 시각화

신경 근육 접합 : 측정 시냅스 크기, 공 초점 형광 현미경을 사용하여 시냅스 단백질 농도의 분열 및 변경

요약

더 많은 비디오 탐색

Quantifying 시냅스 : 시냅스 번호를 수치 Immunocytochemistry 기반 어세이

교양 CNS의 뉴런의 돌기 스파인 형태론 분석

에서 시냅틱 형광의 자동 부량

DetectSyn: Synapse 밀도의 변화를 감지하는 빠르고 편향되지 않은 형광 방법

높은 처리량 및 다중 파라 메트릭 정량화를위한 ​​두 알고리즘

면역형광및 전체산 신경근육 접합의 형태 분석에 대한 단일 골격 근육 섬유의 해부

근위축성 측삭 경화증 모델에서 시냅스 기능의 실시간 형광 측정

결합된 공초점 및 초분해능 현미경에 의한 마우스의 신경근 접합부 특성 분석

Registration of Calcium Transients in Mouse Neuromuscular Junction with High Temporal Resolution using Confocal Microscopy

쥐 내측 위장근에서 신경근 접합부의 형태학적 특성 시각화

높은 처리량 및 다중 파라 메트릭 정량화를위한 두 알고리즘