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Visualization of Germinosomes and the Inner Membrane in Bacillus subtilis Spores
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Chapters
- 00:04Title
- 00:34High Precision Coverslip and Glass Slide Preparation
- 01:17Fluorescent Microsphere and Spore Sample Mounting
- 02:32Fluorescent Microsphere and Spore Imaging
- 03:38FM4-64 Stained PS4150 Spore 3D-SIM Raw Image Reconstruction
- 05:00Image Analysis
- 06:25Results: Representative Germinosome and Inner Membrane Visualization
- 08:04Conclusion
Germinant receptor proteins cluster in ‘germinosomes’ in the inner membrane of Bacillus subtilis spores. We describe a protocol using super resolution microscopy and fluorescent reporter proteins to visualize germinosomes. The protocol also identifies spore inner membrane domains that are preferentially stained with the membrane dye FM4-64.
Tags
Keywords:
GerminosomesInner MembraneBacillus Subtilis SporesSuper-resolution Structured Illumination Microscopy3D-SIMFluorescent Reporter ProteinsSpore MembranesLipid DomainsCoverslipsMicroscope SlidesFluorescent MicrospheresAgaroseUltrapure WaterGene FrameStructured Illumination MicroscopePoint Spread FunctionExcitation Wavelengths
