Optical spectral analysis and multispectral imaging provide powerful means for characterizing samples in a wide variety of applications and on many spatial scales. We present a simple implementation of these techniques in the context of microscopy. A modified commercial microscope equipped with a CMOS imaging detector, combined with an array of light emitting diodes with emission ranging from ultraviolet to near-infrared wavelengths, is described, and examples of information enhancement using multivariate analysis are presented.
REFERENCES
1.
R. C.
Gonzalez
and R. E.
Woods
, Digital Image Processing
, 3rd ed. (Pearson Prentice Hall
, Upper Saddle River, NJ
, 2008
).2.
S.
Svanberg
, Multispectral Imaging-From Astronomy to Microscopy-From Radiowaves to Gammarays
(Springer Verlag
, Berlin
, 2009
).3.
A. P.
Cracknell
and L. W. B.
Hayes
, Introduction to Remote Sensing
, 2nd ed. (CRC
, Boca Raton, FL
, 2007
).4.
M.
Borengasser
, W. S.
Hungate
, and R.
Watkins
, Hyperspectral Remote Sensing: Principles and Applications
(CRC
, Boca Raton, FL
, 2008
).5.
A. K.
Maini
and V.
Agrawal
, Satellite Technology: Principles and Applications
(Wiley
, Chichester
, 2007
).6.
7.
D. B.
Murphy
, Fundamentals of Light Microscopy and Electronic Imaging
(Wiley-Liss
, New York
, 2001
).8.
F. J.
Kao
and P.
Török
, Optical Imaging and Microscopy: Techniques and Advanced Systems
, 2nd ed. (Springer Verlag
, Berlin
, 2007
).9.
J. R.
Lakowicz
, Principles of Fluorescence Spectroscopy
, 3rd ed. (Springer
, New York
, 2006
).10.
T. W.
Anderson
, An Introduction to Multivariate Statistical Analysis
, 3rd ed. (Wiley
, Hoboken, NJ
, 2003
).11.
12.
K. R.
Beebe
and B. R.
Kowalski
, “An introduction to multivariate calibration and analysis
,” Anal. Chem.
59
, 1007A
–1017A
(1987
).13.
P.
Weibring
, T.
Johansson
, H.
Edner
, S.
Svanberg
, B.
Sundnér
, V.
Raimondi
, G.
Cecchi
, and L.
Pantani
, “Fluorescence lidar imaging of historical monuments
,” Appl. Opt.
40
, 6111
–6120
(2001
);
[PubMed]
ibid.
P.
Weibring
, T.
Johansson
, H.
Edner
, S.
Svanberg
, B.
Sundnér
, V.
Raimondi
, G.
Cecchi
, and L.
Pantani
, Appl. Opt.
41
, 434
–436
(2002
).14.
MATLAB, Help files, The Mathworks.
15.
An early application of the use of LEDs in spectroscopy is demonstrated in
P. H.
Hauser
, T. W. T.
Rupasinghe
, and N. E.
Cates
, “A multi-wavelength photometer based on light-emitting diodes
,” Talanta
42
, 605
–612
(1995
).16.
G. C.
Holst
and T. S.
Lomheim
, CMOS/CCD Sensors and Camera Systems
(JCD Publishing, SPIE
, Bellingham, WA
, 2007
).17.
18.
19.
E. J.
Warrant
and D. E.
Nilsson
, Invertebrate Vision
(Cambridge U.P.
, Cambridge
, 2006
).20.
T. H.
Goldsmith
, “What birds see
,” Sci. Am.
295
(1
), 68
–75
(2006
).21.
N. S.
Hart
and M.
Vorobyev
, “Modelling oil droplet absorption spectra and spectral sensitivities of bird cone photoreceptors
,” J. Comp. Physiol., A
191
, 381
–392
(2005
).22.
MATLAB, Image Processing Toolbox, User’s Guide, The Mathworks.
23.
M.
Brydegaard
and S.
Svanberg
, “Contrast functions and spectral data handling
,” in Proceedings, Optics and Laser Applications in Medicine and Enviromental Monitoring for Sustainable Development
, edited by P.
Buah-Bassuah
(University of Cape Coast
, Ghana
, 2007
), pp. 91
–92
.24.
R. R.
Ogle
and M. J.
Fox
, Atlas of Human Hair Microscopic Characteristics
(CRC
, Boca Raton, FL
, 1999
).25.
M.
Brydegaard
, Z. G.
Guan
, and S.
Svanberg
, “Light-emitting diode (LED) based multispectral microscope for imaging transmission spectroscopy
,” Lund Reports on Atomic Physics LRAP-395 (2008
).26.
H.
Moravec
, “When will computer hardware match the human brain?
,” J. Evol. Techn.
1
, 1
–12
(1998
).27.
Roithner LaserTechnik GmbH (Austria), ⟨www.roithner-laser.com⟩.
28.
B.
Standish
, “LEDs for bioanalytical and medical instruments
,” Biophotonics Int.
14
, 37
–39
(2007
).29.
P.
Herman
, B. P.
Maliwal
, H. J.
Lin
, and J. R.
Lakowicz
, “Frequency-domain fluorescence microscopy with the LED as a light source
,” J. Microsc.
203
, 176
–181
(2001
).30.
J. T.
Zoueu
, G. L.
Loum
, T. C.
Haba
, M.
Brydegaard
, and H.
Menan
, J. Appl. Sci.
8
, 2711
–2717
(2008
).© 2009 American Association of Physics Teachers.
2009
American Association of Physics Teachers
AAPT members receive access to the American Journal of Physics and The Physics Teacher as a member benefit. To learn more about this member benefit and becoming an AAPT member, visit the Joining AAPT page.
