TY - GEN
T1 - Simplified approach makes two-photon Multicolor imaging less costly
AU - Franke, Tilman
AU - Serrano-Velez, Jose L.
AU - Rivera-Rivera, Nydia L.
AU - Rosa-Molinar, Eduardo
AU - Langhorst, Matthias
PY - 2014/2
Y1 - 2014/2
N2 - A simplified approach to two-photon multicolor imaging (TPM) is proposed. The simplified multicolor TPM is based on an FEI Intravital2P microscope equipped with a Toptica FemtoFErb 780 fiber laser with 50-mW average power, 100-fs pulses and 100-MHz repetition rate. The laser is passively cooled and small enough to be easily attached to the microscope frame. The detection system is based on a pair of GaAsP PMT detectors. The 562-nm long-pass beamsplitter slices the spectrum into two color channels: blue/green and red. Scanner and PMTs are optimized for high-efficiency dual-color recordings. A hyperspectral detection system that can be plugged in in place of one of the PMTs is designed. Fiji, a life-science-oriented distribution of ImageJ, was used to generate sum spectra and pseudo-real-color representations, and the PoissonNMF plug-in for blind spectral unmixing. Using the simplified TPM, entire whole- mount samples with laser power at 20 mW or lower was imaged.
AB - A simplified approach to two-photon multicolor imaging (TPM) is proposed. The simplified multicolor TPM is based on an FEI Intravital2P microscope equipped with a Toptica FemtoFErb 780 fiber laser with 50-mW average power, 100-fs pulses and 100-MHz repetition rate. The laser is passively cooled and small enough to be easily attached to the microscope frame. The detection system is based on a pair of GaAsP PMT detectors. The 562-nm long-pass beamsplitter slices the spectrum into two color channels: blue/green and red. Scanner and PMTs are optimized for high-efficiency dual-color recordings. A hyperspectral detection system that can be plugged in in place of one of the PMTs is designed. Fiji, a life-science-oriented distribution of ImageJ, was used to generate sum spectra and pseudo-real-color representations, and the PoissonNMF plug-in for blind spectral unmixing. Using the simplified TPM, entire whole- mount samples with laser power at 20 mW or lower was imaged.
UR - https://www.scopus.com/pages/publications/84896950075
M3 - Article
AN - SCOPUS:84896950075
SN - 1081-8693
VL - 21
SP - 20
EP - 23
JO - Biophotonics International
JF - Biophotonics International
ER -