Abstract
In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry. In this study, we enhanced the collected luminescence signal by the use of alternating multiply-coated layers of tantalum pentoxide (Ta2O5) and silicon dioxide (SiO2) on standard microscope cover glasses that has high transmission in the near-infrared wavelength region and high reflection of the visible, luminescence signal. Our coating is biocompatible, allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells. In all cases, the use of coated cover glass enhanced signal, optimally by a factor of about 2. Image analysis of labeled 3T3 cells also shows signal enhancement did not contribute to additional photobleaching. Our results show that properly designed coated cover glass can enhance detected signal in multiphoton microscopy and result in improved image quality.
Highlights
As in most forms of optical microscopic imaging, multiphoton microscopy relies on the use of glass substrates as the viewing window of biological specimens
In nonlinear optical imaging of biological specimens, more than half of the generated luminescence signal is lost, when signal collection is performed in the epi-illuminated geometry
Allows visual examination of the specimens and optimize collection of the luminescence signal. We demonstrated this approach on a number of specimens including sulforhodamine solution, fluorescence microspheres, and labeled 3T3 cells
Summary
As in most forms of optical microscopic imaging, multiphoton microscopy relies on the use of glass substrates as the viewing window of biological specimens. The invention and subsequent development of multiphoton microscopy have demonstrated that this technique is ideal for imaging three-dimensional tissue specimens [1,2]. Our coating is designed for the optimization of reflected luminescence signal while allowing transmission of the near-infrared excitation source. The coating we use allows standard white light examination of the specimens to be achieved. The transmission characteristics of the coated cover glasses conveniently allow standard white light examination of the specimens to be achieved
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
