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Light-emitting diode flashlights as effective and inexpensive light sources for fluorescence microscopy.

ABSTRACT: Light-emitting diodes (LEDs) are becoming more commonly used as light sources for fluorescence microscopy. We describe the adaptation of a commercially available light-emitting diode flashlight for use as a source for fluorescence excitation. This light source is long-lived, inexpensive and is effective for excitation in the range of 440-600 nm.

SUBMITTER: Robertson JB

PROVIDER: S-EPMC2751867 | biostudies-literature | 2009 Oct

REPOSITORIES: biostudies-literature

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Light-emitting diode flashlights as effective and inexpensive light sources for fluorescence microscopy.

Robertson J B JB Zhang Y Y Johnson C H CH

Journal of microscopy 20091001 1

Light-emitting diodes (LEDs) are becoming more commonly used as light sources for fluorescence microscopy. We describe the adaptation of a commercially available light-emitting diode flashlight for use as a source for fluorescence excitation. This light source is long-lived, inexpensive and is effective for excitation in the range of 440-600 nm. ...[more]

PMID: 19772530

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Project description:Chlorophyll fluorescence measured at the leaf scale through pulse amplitude modulation (PAM) has provided valuable insight into photosynthesis. At the canopy- and satellite-scale, solar-induced fluorescence (SIF) provides a method to estimate the photosynthetic activity of plants across spatiotemporal scales. However, retrieving SIF signal remotely requires instruments with high spectral resolution, making it difficult and often expensive to measure canopy-level steady-state chlorophyll fluorescence under natural sunlight. Considering this, we built a novel low-cost photodiode system that retrieves far-red chlorophyll fluorescence emission induced by a blue light emitting diode (LED) light source, for 2 h at night, above the canopy. Our objective was to determine if an active remote sensing-based night-time photodiode method could track changes in canopy-scale LED-induced chlorophyll fluorescence (LEDIF) during an imposed drought on a broadleaf evergreen shrub, Polygala myrtifolia. Far-red LEDIF (720-740 nm) was retrieved using low-cost photodiodes (LEDIFphotodiode) and validated against measurements from a hyperspectral spectroradiometer (LEDIFhyperspectral). To link the LEDIF signal with physiological drought response, we tracked stomatal conductance (gsw) using a porometer, two leaf-level vegetation indices-photochemical reflectance index and normalized difference vegetation index-to represent xanthophyll and chlorophyll pigment dynamics, respectively, and a PAM fluorimeter to measure photochemical and non-photochemical dynamics. Our results demonstrate a similar performance between the photodiode and hyperspectral retrievals of LEDIF (R2 = 0.77). Furthermore, LEDIFphotodiode closely tracked drought responses associated with a decrease in photochemical quenching (R2 = 0.69), Fv/Fm (R2 = 0.59) and leaf-level photochemical reflectance index (R2 = 0.59). Therefore, the low-cost LEDIFphotodiode approach has the potential to be a meaningful indicator of photosynthetic activity at spatial scales greater than an individual leaf and over time.

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Light-emitting diode flashlights as effective and inexpensive light sources for fluorescence microscopy.

Publications

Light-emitting diode flashlights as effective and inexpensive light sources for fluorescence microscopy.

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