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Updated: Jul 4, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Fluorescence imaging using a fluorescent protein with a large Stokes shift.

Takako Kogure¹, Hiroyuki Kawano, Yukiko Abe

¹Laboratory for Cell Function and Dynamics, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama 351-0198, Japan.

Methods (San Diego, Calif.)

|July 1, 2008

Summary

Keima, a far-red fluorescent protein, offers a large Stokes shift for dual-color imaging. Its variants, including tandem dimer Keima (tdKeima), enhance fluorescence cross-correlation spectroscopy and two-photon laser scanning microscopy applications.

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Area of Science:

Biophysics
Molecular Biology
Microscopy

Background:

Keima is a far-red fluorescent protein with a large Stokes shift, absorbing at ~440nm and emitting at ~620nm.
Tetrameric (tKeima), dimeric (dKeima), and monomeric (mKeima) versions exist, with tandem dimer Keima (tdKeima) being the brightest.

Purpose of the Study:

To demonstrate the utility of Keima variants in dual-color fluorescence imaging.
To showcase applications in fluorescence cross-correlation spectroscopy (FCCS) and two-photon laser scanning microscopy (TPLSM).

Main Methods:

Utilizing Keima's large Stokes shift for simultaneous excitation with other fluorescent proteins.
Employing Keima in conjunction with fluorescent proteins possessing smaller Stokes shifts.

Main Results:

Keima enables effective dual-color imaging when paired with fluorescent proteins of smaller Stokes shifts.
Demonstrated successful application in advanced imaging techniques like FCCS and TPLSM.

Conclusions:

Keima and its variants are valuable tools for advanced dual-color fluorescence imaging.
The large Stokes shift of Keima facilitates multicolor imaging strategies, improving resolution and data acquisition.