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. 2007 Apr 1;3(4):251-6.
doi: 10.7150/ijbs.3.251.

A polychromator-based microspectrophotometer

Valter Evangelista  1 , Mauro Evangelisti, Laura Barsanti, Anna Maria Frassanito, Vincenzo Passarelli, Paolo Gualtieri
Affiliations

A polychromator-based microspectrophotometer

Valter Evangelista et al. Int J Biol Sci. .

Abstract

A microspectrophotometer is a digital microscope used to measure absorption and fluorescence spectra. In this paper we describe a polychromator-based microspectrophotometer that performs in vivo absorption or emission measurements at the same time on different subcellular compartments such as photoreceptive and photosynthetic structures of algal cells. In this system, a flat field imaging concave grating polychromator is connected to the slit-shaped exit pupil of a light-guide probe mounted onto a microscope equipped with an epifluorescence module. The subcellular components, on which the spectra will be measured, are placed in the microscope field and finely adjusted. The outer bundle of the probe is used for centering the objects, while the central bundle of the probe, containing 19 light guides, is used for acquiring either transmitted or emitted light (i.e. fluorescence). The light transmitted or emitted by the subcellular components is collected by the probe mounted in the back focal plane of the ocular. The exit pupil of this probe, connected to a flat field imaging concave grating polychromator, produces a dispersion image that in turn is focused onto a digital slow scan cooled CCD camera. Absorption and emission spectra of algal subcellular compartments are presented.

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Conflict of interest statement

Conflict of interest: The authors have declared that no conflict of interest exists.

Figures

Figure 1
Figure 1
Set-up of the polychromator-based microspectrophotometer
Figure 2
Figure 2
Entrance (a) and exit (b) pupils of the light guide probe
Figure 3
Figure 3
Layout of the software interface
Figure 4
Figure 4
a) Bright field image of Dunaliella. b) Position of the probe upon Dunaliella
Figure 4
Figure 4
a) Bright field image of Dunaliella. b) Position of the probe upon Dunaliella
Figure 5
Figure 5
Absorption spectra of the eyespot (a) and a chloroplast (b) of Dunaliella
Figure 5
Figure 5
Absorption spectra of the eyespot (a) and a chloroplast (b) of Dunaliella
Figure 6
Figure 6
a) Fluorescent image of Euglena gracilis; b) Position of the probe upon Euglena gracilis
Figure 6
Figure 6
a) Fluorescent image of Euglena gracilis; b) Position of the probe upon Euglena gracilis
Figure 7
Figure 7
Emission spectra of the photoreceptor (a) and a chloroplast (b) of Euglena gracilis
Figure 7
Figure 7
Emission spectra of the photoreceptor (a) and a chloroplast (b) of Euglena gracilis
See this image and copyright information in PMC

References

    1. Barsanti L, Gualtieri P. Algae, Anatomy, Biochemistry, and Biotechnology. Boca Raton: Taylor & Francis; 2005.
    1. Piller H. Microscope photometry. Berlin: Springer-Verlag; 1977.
    1. Gualtieri P. Microspectroscopy of photoreceptor pigments in flagellated algae. Critical Rev. Plant Sci. 1991;9(6):474–495.
    1. Millie D, Evens TJ. Using absorbance and fluorescence spectra to discriminate microalgae. Eur. J. Phycol. 2002;37:313–322.
    1. Cramer M, Myers J. Growth and photosynthetic characteristics of Euglena gracilis. Arch Mikrobiol. 1952;17:384–402.

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