Abstract
Techniques used to clear biological tissue for fluorescence microscopy are essential to connect anatomical principles at levels ranging from subcellular to the whole animal. Here we report a simple and straightforward approach to efficiently render opaque tissue samples transparent and show that this approach can be modified to rapidly label intact tissue samples with antibodies for large volume fluorescence microscopy. This strategy applies a magnetohydrodynamic (MHD) force to accelerate the removal of lipids from tissue samples at least as large as an intact adult mouse brain. We also show that MHD force can be used to accelerate antibody penetration into tissue samples. This strategy complements a growing array of tools that enable high-resolution 3-dimensional anatomical analyses in intact tissues using fluorescence microscopy. MHD-accelerated clearing is simple, fast, reliable, inexpensive, provides good thermal regulation, and is compatible with existing strategies for high-quality fluorescence microscopy of intact tissues.
Highlights
Techniques used to clear biological tissue for fluorescence microscopy are essential to connect anatomical principles at levels ranging from subcellular to the whole animal
High-quality images can be produced with both Electric-only and MHD-accelerated clearing, and no differences in shape were observed for tissue samples prepared with MHD-accelerated versus electric-only clearing; samples prepared with MHD-accelerated clearing (30 VDC; 0.35 AMPS) were ready to be imaged in less time than electric-only clearing (30 VDC; 0.35 AMPS) and exhibited better imaging (Fig. S2)
MHD accelerated clearing maintains all advantages of electric-only clearing and adds the additional MHD force to further accelerate tissue clearing without increasing the potentially damaging electric field (Table 1,Fig. 2; Fig. S1; Fig. S2)
Summary
Techniques used to clear biological tissue for fluorescence microscopy are essential to connect anatomical principles at levels ranging from subcellular to the whole animal. We report a simple and straightforward approach to efficiently render opaque tissue samples transparent and show that this approach can be modified to rapidly label intact tissue samples with antibodies for large volume fluorescence microscopy. This strategy applies a magnetohydrodynamic (MHD) force to accelerate the removal of lipids from tissue samples at least as large as an intact adult mouse brain. We show that MHD force can be used to accelerate antibody penetration into tissue samples This strategy complements a growing array of tools that enable high-resolution 3-dimensional anatomical analyses in intact tissues using fluorescence microscopy. These MHD-based approaches work in both vertebrate (shown for mouse and zebrafish) and invertebrate
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