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Information Journal Paper

Title

SKELETAL MUSCLE CLARITY: A PRELIMINARY STUDY OF IMAGING THE THREE-DIMENSIONAL ARCHITECTURE OF BLOOD VESSELS AND NEURONS

Pages

  132-137

Keywords

Abstract

 Objective: Passive CLARITY is a whole-tissue clearing protocol, based on sodium dodecyl sulfate (SDS) clearing, for imaging intact tissue containing transgenic or immunolabeled fluorescent proteins. In this study, we present an improved passive CLARITY protocol with efficient immunolabeling without the need for electrophoresis or complex instrumentation.Materials and Methods: In this experimental study, after perfusion of C57BL/6N mice with phosphate-buffered saline (PBS) and then with acrylamide-paraformaldehyde (PFA), the quadriceps femoris MUSCLE was removed. The MUSCLE samples were post-fixed and degassed to initiate polymerization. After removing the excess hydrogel around the MUSCLE, lipids were washed out with the passive CLARITY technique. The transparent whole intact MUSCLEs were labeled for VESSEL and NEURON markers, and then imaged by confocal microscopy. Three-dimensional images were reconstructed to present the MUSCLE tissue architecture. Results: We established a simple clearing protocol using wild type MOUSE MUSCLE and labeling of vasculatures and NEURONs. Imaging the fluorescent signal was achieved by protein fixation, adjusting the pH of the SDS solution and using an optimum temperature (37oC) for tissue clearing, all of which contributed to the superiority of our protocol. Conclusion: We conclude that this passive CLARITY protocol can be successfully applied to three-dimensional cellular and whole MUSCLE imaging in mice, and will facilitate structural analyses and connectomics of large assemblies of MUSCLE cells, VESSELs and NEURONs in the context of three-dimensional systems.

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    Cite

    APA: Copy

    ZHANG, WENLI, LIU, SHAOHUA, ZHANG, WEICHEN, HU, WEI, JIANG, MIN, TAMADON, AMIN, & FENG, YI.. (2018). SKELETAL MUSCLE CLARITY: A PRELIMINARY STUDY OF IMAGING THE THREE-DIMENSIONAL ARCHITECTURE OF BLOOD VESSELS AND NEURONS. CELL JOURNAL (YAKHTEH), 20(2), 132-137. SID. https://sid.ir/paper/712192/en

    Vancouver: Copy

    ZHANG WENLI, LIU SHAOHUA, ZHANG WEICHEN, HU WEI, JIANG MIN, TAMADON AMIN, FENG YI.. SKELETAL MUSCLE CLARITY: A PRELIMINARY STUDY OF IMAGING THE THREE-DIMENSIONAL ARCHITECTURE OF BLOOD VESSELS AND NEURONS. CELL JOURNAL (YAKHTEH)[Internet]. 2018;20(2):132-137. Available from: https://sid.ir/paper/712192/en

    IEEE: Copy

    WENLI ZHANG, SHAOHUA LIU, WEICHEN ZHANG, WEI HU, MIN JIANG, AMIN TAMADON, and YI. FENG, “SKELETAL MUSCLE CLARITY: A PRELIMINARY STUDY OF IMAGING THE THREE-DIMENSIONAL ARCHITECTURE OF BLOOD VESSELS AND NEURONS,” CELL JOURNAL (YAKHTEH), vol. 20, no. 2, pp. 132–137, 2018, [Online]. Available: https://sid.ir/paper/712192/en

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