Laboratory Soft X-Ray Microscopy with an Integrated Visible-Light Microscope-Correlative Workflow for Faster 3D Cell...

Area of Science:

• Biomedical imaging
• Microscopy techniques
• Cell biology

Background:

• Laboratory transmission soft X-ray microscopy (L-TXM) offers high-resolution 3D imaging but faces limitations.
• Key challenges include a small field of view and potential rotation stage misalignment.
• These issues hinder efficient sample navigation and tomographic data acquisition.

Purpose of the Study:

• To address operational challenges in L-TXM.
• To integrate a visible-light microscope (VLM) into the L-TXM system.
• To improve sample navigation and workflow efficiency for 3D imaging.

Main Methods:

• Developed a calibration procedure for the goniometer sample stage and integrated VLM.
• Utilized the VLM for pre-experiment sample navigation and alignment.
• Performed 3D imaging of a test sample and THP-1 cell extracellular matrix.

Main Results:

• Successful integration of VLM into L-TXM system overcame field of view and alignment issues.
• Demonstrated improved sample navigation and efficient tomographic data acquisition.
• Achieved novel visualization of THP-1 cell extracellular matrix using L-TXM.

Conclusions:

• Integrated VLM effectively resolves major L-TXM operational limitations.
• The enhanced L-TXM system facilitates high-resolution 3D imaging of biological samples.
• This advancement supports ongoing biomedical research, particularly in cell biology.