Myosin rods are a source of second harmonic generation signals in skeletal muscle Export

@inproceedings{citeulike:6088342, abstract = {Intrinsic second harmonic generation (SHG) signals can be used to visualize the three-dimensional structure of cardiac and skeletal muscle with high spatial resolution. Fluorescence labeling of complementary sarcomeric proteins, e.g. actin, indicates that the observed SHG signals arise from the myosin filaments. Recently, the myosin rod domain or LMM - light meromyosin - has been reported to be the dominant source of this SHG signal. However, to date, mostly negative and indirect evidence has been presented to support this assumption. Here, we show, to our knowledge, the first direct evidences that strong SHG signals can be obtained from synthetic paracrystals. These rod shaped filaments are formed from purified LMM. SDS-PAGE protein analysis confirmed that the LMM crystals lack myosin head domains. Some regions of the LMM paracrystals produce a strong SHG signal whereas others did not. The SHG signals were recorded with a laser-scanning microscope (Leica SP2). A ps laser tuned to 880 nm was used to excite the sample through an 63x objective of 1.2 NA. In order to visualize the synthetic filaments - in addition to SHG imaging -, the LMM was labeled with the fluorescent marker 5-IAF. We were able to observe filaments of 1 to 50 \µm in length and of up to 5 \µm in diameter. In conclusion, we can show that the myosin rod domain (LMM) is a dominant source for intrinsic SHG signals. There seems, however, a signal dependence on the paracrystals' morphology. This dependence is being investigated.}, author = {Sch\"{u}rmann, Sebastian and Weber, Cornelia and Fink, Rainer H. A. and Vogel, Martin}, citeulike-article-id = {6088342}, citeulike-linkout-0 = {http://scitation.aip.org/getabs/servlet/GetabsServlet?prog=normal&id=PSISDG00644200000164421U000001&idtype=cvips&gifs=yes}, citeulike-linkout-1 = {http://link.aip.org/link/?PSI/6442/64421U}, editor = {Periasamy, Ammasi and So, Peter T. C.}, journal = {Multiphoton Microscopy in the Biomedical Sciences VII}, keywords = {myosin, shg}, location = {San Jose, CA, USA}, number = {1}, pages = {64421U+}, posted-at = {2009-11-09 13:06:34}, priority = {2}, publisher = {SPIE}, title = {Myosin rods are a source of second harmonic generation signals in skeletal muscle}, url = {http://scitation.aip.org/getabs/servlet/GetabsServlet?prog=normal&id=PSISDG00644200000164421U000001&idtype=cvips&gifs=yes}, volume = {6442}, year = {2007} }

TY - CONF ID - citeulike:6088342 L3 - citeulike-article-id:6088342 N2 - Intrinsic second harmonic generation (SHG) signals can be used to visualize the three-dimensional structure of cardiac and skeletal muscle with high spatial resolution. Fluorescence labeling of complementary sarcomeric proteins, e.g. actin, indicates that the observed SHG signals arise from the myosin filaments. Recently, the myosin rod domain or LMM - light meromyosin - has been reported to be the dominant source of this SHG signal. However, to date, mostly negative and indirect evidence has been presented to support this assumption. Here, we show, to our knowledge, the first direct evidences that strong SHG signals can be obtained from synthetic paracrystals. These rod shaped filaments are formed from purified LMM. SDS-PAGE protein analysis confirmed that the LMM crystals lack myosin head domains. Some regions of the LMM paracrystals produce a strong SHG signal whereas others did not. The SHG signals were recorded with a laser-scanning microscope (Leica SP2). A ps laser tuned to 880 nm was used to excite the sample through an 63x objective of 1.2 NA. In order to visualize the synthetic filaments - in addition to SHG imaging -, the LMM was labeled with the fluorescent marker 5-IAF. We were able to observe filaments of 1 to 50 µm in length and of up to 5 µm in diameter. In conclusion, we can show that the myosin rod domain (LMM) is a dominant source for intrinsic SHG signals. There seems, however, a signal dependence on the paracrystals' morphology. This dependence is being investigated. CY - San Jose, CA, USA IS - 1 JF - Multiphoton Microscopy in the Biomedical Sciences VII TI - Myosin rods are a source of second harmonic generation signals in skeletal muscle PB - SPIE VL - 6442 SP - 64421U KW - myosin KW - shg AU - Schürmann, Sebastian AU - Weber, Cornelia AU - Fink, Rainer AU - Vogel, Martin A2 - Periasamy, Ammasi A2 - So, Peter PY - 2007/// UR - http://scitation.aip.org/getabs/servlet/GetabsServlet?prog=normal&id=PSISDG00644200000164421U000001&idtype=cvips&gifs=yes ER -