Time-resolved two-photon excitation fluorescence spectroscopy and microscopy using a high repetition rate streak camera

2007 ◽  
Vol 3 (2) ◽  
pp. 157-160
Author(s):  
Li-xin Liu ◽  
Ju-le Qu ◽  
Zi-yang Lin ◽  
Lei Wang ◽  
Zhe Fu ◽  
...  
Keyword(s):  
Fluorescence Spectroscopy ◽  
Repetition Rate ◽  
Streak Camera ◽  
High Repetition Rate ◽  
Time Resolved ◽  
Two Photon ◽  
High Repetition ◽  
Photon Excitation ◽  
Two Photon Excitation

Time-resolved Two-photon Excitation Fluorescence Spectroscopy Using a Streak Camera

2007 ◽  
Author(s):  
Lixin Liu ◽  
Junle Qu ◽  
Ziyang Lin ◽  
Baoping Guo ◽  
Wenqing Liu ◽  
...  
Keyword(s):  
Fluorescence Spectroscopy ◽  
Streak Camera ◽  
Time Resolved ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

scholarly journals Time-resolved two-photon excitation fluorescence spectroscopy

2006 ◽  
Vol 55 (12) ◽  
pp. 6281
Author(s):  
Liu Li-Xin ◽  
Qu Jun-Le ◽  
Lin Zi-Yang ◽  
Chen Dan-Ni ◽  
Xu Gai-Xia ◽  
...  
Keyword(s):  
Fluorescence Spectroscopy ◽  
Time Resolved ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

Simultaneous, time-resolved measurements of OH and HO2 radicals by coupling of high repetition rate LIF and cw-CRDS techniques to a laser photolysis reactor and its application to the photolysis of H2O2

2010 ◽  
Vol 103 (3) ◽  
pp. 725-733 ◽  
Author(s):  
A. E. Parker ◽  
C. Jain ◽  
C. Schoemaecker ◽  
P. Szriftgiser ◽  
O. Votava ◽  
...  
Keyword(s):  
Repetition Rate ◽  
High Repetition Rate ◽  
Laser Photolysis ◽  
Time Resolved ◽  
High Repetition ◽  
Time Resolved Measurements

scholarly journals Improving Signal and Photobleaching Characteristics of Temporal Focusing Microscopy with the Increase in Pulse Repetition Rate

2019 ◽  
Vol 2 (3) ◽  
pp. 65
Author(s):  
Viktoras Lisicovas ◽  
Bala Murali Krishna Mariserla ◽  
Chakradhar Sahoo ◽  
Reuben T. Harding ◽  
Michael K. L. Man ◽  
...  
Keyword(s):  
Pulse Energy ◽  
Repetition Rate ◽  
Signal Intensity ◽  
High Repetition Rate ◽  
High Peak Power ◽  
High Pulse Energy ◽  
Two Photon ◽  
High Repetition ◽  
Laser Systems ◽  
High Pulse

Wide-field temporal focused (WF-TeFo) two-photon microscopy allows for the simultaneous imaging of a large planar area, with a potential order of magnitude enhancement in the speed of volumetric imaging. To date, low repetition rate laser sources with over half a millijoule per pulse have been required in order to provide the high peak power densities for effective two-photon excitation over the large area. However, this configuration suffers from reduced signal intensity due to the low repetition rate, saturation effects due to increased excitation fluences, as well as faster photobleaching of the fluorescence probe. In contrast, with the recent advent of high repetition rate, high pulse energy laser systems could potentially provide the advantages of high repetition rate systems that are seen in traditional two-photon microscopes, while minimizing the negatives of high fluences in WF-TeFo setups to date. Here, we use a 100 microjoule/high repetition rate (50–100 kHz) laser system to investigate the performance of a WF-TeFo two-photon microscope. While using micro-beads as a sample, we demonstrate a proportionate increase in signal intensity with repetition rate, at no added cost in photobleaching. By decreasing pulse intensity, via a corresponding increase in repetition rate to maintain fluorescence signal intensity, we find that the photobleaching rate is reduced by ~98.4%. We then image live C. elegans at a high repetition rate for 25 min. as a proof-of-principle. Lastly, we identify the steady state temperature increase as the limiting process in further increasing the repetition rate, and we estimate that repetition rate in the range between 0.5 and 5 MHz is ideal for live imaging with a simple theoretical model. With new generation low-cost fiber laser systems offering high pulse energy/high repetition rates in what is essentially a turn-key solution, we anticipate increased adoption of this microscopy technique by the neuroscience community.

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