Economic and simple system to combine single-spot photolysis and whole-field fluorescence imaging. - Archive ouverte HAL Access content directly
Journal Articles Journal of Biomedical Optics Year : 2013

Economic and simple system to combine single-spot photolysis and whole-field fluorescence imaging.

(1, 2) , (3) , (3) , (4)
1
2
3
4

Abstract

ABSTRACT. In recent years, the use of light emitting diodes (LEDs) has become commonplace in fluorescence microscopy. LEDs are economical and easy to couple to commercial microscopes, and they provide powerful and stable light that can be triggered by transistor-transistor logic pulses in the range of tens of microseconds or shorter. LEDs are usually installed on the epifluorescence port of the microscope to obtain whole-field illumination, which is ideal for fluorescence imaging. In contrast, photolysis or channelrhodopsin stimulation often requires localized illumination, typically achieved using lasers. Here we show that insertion of a long-pass (>411 nm) filter with an appropriately sized pinhole in the epifluorescence pathway, combined with dual UV/visible illumination, can produce efficient whole-field visible illumination and spot UV illumination of 15 to 20 μm. We tested our system by performing calcium imaging experiments combined with L-glutamate or N-methyl-D-aspartic acid (NMDA) photorelease in hippocampal neurons from brain slices or dissociated cultures, demonstrating the ability to obtain local activation of NMDA receptors exclusively in the illuminated spot. The very inexpensive and simple system that we report here will allow many laboratories with limited budgets to run similar experiments in a variety of physiological applications.
Fichier principal
Vignette du fichier
Jaafari_2013_Economic_MA.pdf (321.91 Ko) Télécharger le fichier
Vignette du fichier
inserm-00843337_edited.pdf (282.2 Ko) Télécharger le fichier
Origin : Files produced by the author(s)
Origin : Files produced by the author(s)
Loading...

Dates and versions

inserm-00843337 , version 1 (11-07-2013)

Identifiers

Cite

Nadia Jaafari, Mark Henson, Jeremy Graham, Marco Canepari. Economic and simple system to combine single-spot photolysis and whole-field fluorescence imaging.. Journal of Biomedical Optics, 2013, 18 (6), pp.60505. ⟨10.1117/1.JBO.18.6.060505⟩. ⟨inserm-00843337⟩
211 View
549 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More