Laboratory based correlative cryo-soft X-ray tomography and cryo-fluorescence microscopy

Area of Science:

• Cell Biology
• Microscopy
• Biophysics

Background:

• Cryo-soft X-ray tomography (CSXT) offers high-resolution, label-free 3D imaging of intact cells.
• Traditional CSXT requires access to specialized synchrotron facilities, limiting its widespread use.
• Correlating CSXT with cryo-fluorescence microscopy enables localization of molecular markers within cellular structures.

Purpose of the Study:

• To introduce a compact, laboratory-based soft X-ray tomography (SXT) microscope, the SXT-100.
• To demonstrate the capability of the SXT-100 for high-throughput, high-resolution cellular imaging.
• To highlight the integration of cryo-fluorescence imaging with the SXT-100 for correlative studies.

Main Methods:

• Utilized fast-freezing for sample preparation, enabling imaging of cryo-preserved cells.
• Employed the compact SXT-100 microscope for soft X-ray tomography.
• Integrated cryo-fluorescence microscopy for correlative imaging of fluorescently tagged cellular components.

Main Results:

• The SXT-100 successfully imaged cryo-prepared samples, demonstrating its utility in disease and drug research.
• The compact system provides high throughput and spatial resolution, comparable to synchrotron-based methods.
• Integrated cryo-fluorescence capability allowed co-localization of fluorescent signals with 3D cellular structures.

Conclusions:

• The SXT-100 significantly increases accessibility to advanced cryo-soft X-ray tomography.
• This technology accelerates research in complex 3D organelle structure, nanoparticle uptake, and rare cellular events.
• The laboratory-based SXT-100 facilitates multimodal imaging workflows and enables use in secure facilities like BSL3 labs.