Incoherent color holography lattice light-sheet for subcellular imaging of dynamic structures

Area of Science:

• Biophysics
• Cell Biology
• Neuroscience

Background:

• Cellular functions and subcellular compartments rely on precise colocalization of proteins and lipids.
• Understanding cell signaling is critical for studying neurodegenerative diseases such as Parkinson's, Alzheimer's, and motoneuron diseases.
• Live cell imaging in intact tissue is essential for observing dynamic cellular processes.

Purpose of the Study:

• To explore the advantages of incoherent color holography lattice light-sheet (ICHLLS) for multiwavelength quantitative monitoring of 3D cellular dynamics.
• To investigate the colocalization of fluorescent markers in live cells within intact tissue using ICHLLS.
• To demonstrate the potential of ICHLLS for studying complex cellular functions and disease mechanisms.

Main Methods:

• Utilizing lattice light-sheet imaging principles.
• Implementing an incoherent detection system with actively controlled dual diffractive lenses and phase-shifting.
• Employing sequential multiwavelength excitation and z-galvo scanning with an extended field-of-view.
• Simultaneously recording multidimensional object waves, including intensity, phase, and wavelength information.

Main Results:

• Demonstrated the capability of ICHLLS to image live cells in intact neural tissue.
• Successfully measured the colocalization of fluorescence indicators within live cells.
• Provided a proof-of-concept for the ICHLLS system's configuration and functionality.

Conclusions:

• ICHLLS offers a powerful tool for multiwavelength quantitative monitoring of 3D cellular dynamics in live tissue.
• The system facilitates the investigation of protein and lipid colocalization, essential for cell signaling.
• ICHLLS has significant potential for advancing research in neurodegenerative diseases by enabling detailed study of cellular functions in vivo.