A Practical Solution for 77 K Fluorescence Measurements Based on LED Excitation and CCD Array Detector

PLoS One. 2015 Jul 15;10(7):e0132258. doi: 10.1371/journal.pone.0132258. eCollection 2015.

Abstract

The fluorescence emission spectrum of photosynthetic microorganisms at liquid nitrogen temperature (77 K) provides important insights into the organization of the photosynthetic machinery of bacteria and eukaryotes, which cannot be observed at room temperature. Conventionally, to obtain such spectra, a large and costly table-top fluorometer is required. Recently portable, reliable, and largely maintenance-free instruments have become available that can be utilized to accomplish a wide variety of spectroscopy-based measurements in photosynthesis research. In this report, we show how to build such an instrument in order to record 77K fluorescence spectra. This instrument consists of a low power monochromatic light-emitting diode (LED), and a portable CCD array based spectrometer. The optical components are coupled together using a fiber optic cable, and a custom made housing that also supports a dewar flask. We demonstrate that this instrument facilitates the reliable determination of chlorophyll fluorescence emission spectra for the cyanobacterium Synechocystis sp. PCC 6803, and the green alga Chlamydomonas reinhardtii.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calibration
  • Chlamydomonas / metabolism
  • Fluorescence
  • Optics and Photonics / instrumentation*
  • Pigments, Biological / analysis
  • Reference Standards
  • Signal-To-Noise Ratio
  • Spectrometry, Fluorescence / methods*
  • Synechocystis / metabolism

Substances

  • Pigments, Biological

Grants and funding

Funding was provided through the PhD position awarded to JJL by the Norwegian University of Science and Technology, Trondheim, Norway. The funder had no role in study design, data collection and analysis, discussion to publish, or preparation of the manuscript.