Multicolor lifetime imaging and its application to HIV-1 uptake

Area of Science:

• Biophysics
• Cell Biology
• Microscopy

Background:

• Multicolor imaging with genetically encodable fluorescent proteins is crucial for live cell studies.
• Simultaneous visualization of multiple cellular components and biological processes remains a challenge.

Purpose of the Study:

• To demonstrate simultaneous imaging of nine fluorescent proteins in a single acquisition.
• To visualize the process of human immunodeficiency virus entry into host cells.
• To showcase the potential for advanced multicolor live cell imaging.

Main Methods:

• Utilizing fluorescence lifetime imaging microscopy (FLIM).
• Employing pulsed interleaved excitation with three laser lines.
• Combining FLIM with genetically encodable fluorescent proteins for multicolor imaging.

Main Results:

• Successfully demonstrated simultaneous imaging of nine fluorescent proteins.
• Visualized quadruple-labeled human immunodeficiency viruses on host cells.
• Enabled simultaneous visualization of sub-cellular organelles (mitochondria, cytoskeleton, nucleus) during virus entry.

Conclusions:

• Multicolor fluorescence lifetime imaging offers a powerful strategy for spatio-temporal live cell imaging.
• This technique significantly advances the ability to study complex biological processes like viral infection.
• The method has the potential to image up to nine spectral channels in living cells, opening new avenues in biological research.