A PVAc-Based Benzophenone-8 Filter as an Alternative to Commercially Available Dichroic Filters for Monitoring Calcium Activity in Live Neurons via Fura-2 AM

Author

Blockstein, Lior ; Luk, Collin C. ; Mudraboyina, Anil Kumar ; Syed, Naweed I. ; Yadid-Pecht, Orly

Author_Institution

Dept. of Electr. & Comput. Eng., Univ. of Calgary, Calgary, AB, Canada

Volume

4

Issue

3

fYear

2012

fDate

6/1/2012 12:00:00 AM

Firstpage

1004

Lastpage

1012

Abstract

In this paper, we design, fabricate, and test a PVAc-based benzophenone-8 (PVAcB-8) emission filter to monitor calcium changes in live neurons. We loaded neurons with fura-2 AM, a calcium-sensitive fluorescent dye, which has dual excitation wavelengths at 340 nm, 380 nm with ratiometric emission at 510 nm. To attenuate excitation wavelengths in the UV region and transmit emission light at 510 nm, benzophenone-8 absorption dye was selected as it exhibits high attenuation across 300 to 400 nm. Measurements on the PVAcB-8 filter revealed an attenuation of approximately two and a half orders of magnitude, which is sufficient to collect fluorescence signal from fura-2 AM loaded neurons. The PVAcB-8 filter was compared with a commercially available dichroic filter and proven more efficient in fluorescent imaging.

Keywords

biomedical equipment; biomedical optical imaging; calcium; filters; fluorescence; neurophysiology; polymers; PVAc-based benzophenone-8 filter; PVAcB-8 emission filter; UV region; benzophenone-8 absorption dye; calcium activity; calcium-sensitive fluorescent dye; dichroic filter; dual excitation wavelength; fluorescence signal; fluorescent imaging; fura-2 AM; fura-2 AM loaded neuron; live neuron; wavelength 340 nm to 510 nm; Absorption; Calcium; Fabrication; Fluorescence; Imaging; Neurons; Optical filters; Benzophenone-8; PVAc polymer; absorption; emission filter; fluorescence; fura-2 AM;

fLanguage

English

Journal_Title

Photonics Journal, IEEE

Publisher

ieee

ISSN

1943-0655

Type

jour

DOI

10.1109/JPHOT.2012.2200668

Filename

6203546