was 80% q u e n c h e d at m a x i m u m compression, a value
equal in m a g n i t u d e to the 1:1 sample. If only collisional
quenching was occurring, t h e n one would have e x p e c t e d
the 99:1 s a m p l e to have exhibited the smallest reduction
in the observed fluorescence. T h i s discrepancy indicates
t h a t other :factors are involved which are related to the
degree of m o n o l a y e r compression.
P h o t o b l e a c h i n g of the dye p r o b a b l y played a role in
the observed fluorescent decrease. However, an emission
decline caused solely by decreased lateral diffusion of
photobleac:hed dye molecules seems an unlikely expla-
nation when one considers our previous L - B resultsp
which were essentially free f r o m p h o t o b l e a c h i n g effects
(in spite of the relatively low lateral diffusion exhibited
b y L - B films). In addition, the work p r e s e n t e d here used
an incident laser intensity of 15 m W , as o p p o s e d to the
m u c h higher intensity of 70 m W used in the L - B work.
In a n y event, the m o s t p r o n o u n c e d effect of p h o t o b l e a c h -
ing would have occurred at higher pressures, where the
fluorescence intensity levels off at a m i n i m u m . T h i s level-
ing is p r o b a b l y the result of increased dye concentration
in the condensed phase, c o u n t e r a c t e d b y e x a c e r b a t e d
p h o t o b l e a c h i n g u n d e r a condition of restricted molecular
mobility.
A possible corollary to the collisional quenching and
p h o t o b l e a c h i n g a r g u m e n t s is suggested b y several stud-
ies which s:how the aggregation of cyanine dyes into do-
m a i n s in lipid vesicles. 1° I t is therefore possible that, as
the m o n o l a y e r was compressed, the highly mobile dye
molecules at the a i r / w a t e r interface m a y have favored
aggregation. In t e r m s of fluorescence, the presence of
d o m a i n s m e a n s a radical increase in dye-dye collision
The Use of Refractive Optics for High-Fidelity
Image Transfer in Spatially Resolved
Spectroscopic Measurements
LARRY R. P A D G E T T and P A U L B. F A R N S W O R T H *
Department of Chemistry, Brigham Young University, Provo, Utah 84602
Measurements of emission intensities from spatially inhomogeneous
sources often :require faithful image reproduction at the entrance slit of
a monochromator or spectrograph, or at some intermediate aperture. In
fundamental studies, the spatial variations in intensity contain infor-
mation about the basic nature of the source. For analytical measure-
ments, the spatial resolution is required for the selection of a region in
the source which exhibits the optimum analytical characteristics. Re-
fractive optics; can be used for this type of image transfer, but only if
proper attentJion is given to the minimization or elimination of lens
aberrations. We have developed a simple lens system that allows for the
elimination of chromatic aberration and the control of spherical aber-
ration. Two piano-convex fused-silica lenses with nominal focal lengths
of 300 mm arc mounted on translation stages that are driven in opposite
Received 21 July 1987; revision received 5 November 1987.
* Author to whom correspondence should be sent.
608 Volume 42, Number 4, 1988 ooo3-7o28/ss/42o4-osos,2.oo/o
© 1988 Society for Applied Spectroscopy
f r e q u e n c y a n d a s h a r p reduction in long-range lateral
diffusion, b o t h of which could have h a d the effect of
decreasing the q u a n t u m efficiency of the monolayer b o u n d
fluorophore.
ACKNOWLEDGMENTS
This work was funded by a Biomedical Research Grant from the
Whitaker Foundation (W.M.R.), by NIH Grant HL32132 (W.M.R.),
and by a travel stipend from the United Nations (S.-R.W.). The authors
would like to extend appreciation to their colleagues at the University
of Utah--& N. Herron, J. T. Ives, P. A. Suci, and D. E. Yoshida--for
helpful discussions.
1. O. S. Wolfbeis, Analytische Chimie 345, 387 (1986).
2. T. M. Freeman and W. R. Seitz, Anal. Chem. 50, 1242 (1978).
3. R. E. Benner and R. K. Chang, "Utilization of Optical Fibers in
Remote Inelastic Light Scattering Probes," in Fiber Optics, B.
Bendow and S. S. Mitra, Eds. (Plenum Press, New York, 1979),
pp. 625-639.
4. W. M. Reichert, J. T. Ives, and P. A. Suci, Appl. Spectrosc. 41,
1347 (1987).
5. P. A. Suci and W. M. Reichert, Appl. Spectrosc. 42, 120 (1988).
6. D. M. Jameson, "Fluorescence: Principles, Methodologies, and Ap-
plications," in Fluorescein Hapten: An Immunological Probe, E.
W. Voss, Ed. (CRC Press, Boca Raton, Florida., 1984), pp. 23-48.
7. H. Kuhn, D. Mobius, and H. Bucher, "Spectroscopy of Monolayer
Assemblies," Part 1: "Principles and Applications," Part 2: "Ex-
perimental Procedure," in Physical Methods of Chemistry, Part
IIIB, A. Weissberger and B. W. Rossiter, Eds. (Wiley-Interscience,
New York, 1972), pp. 577-701.
8. J. R. Lokowicz, Principles of Fluorescence Spectroscopy (Plenum
Press, New York, 1983).
9. G. L. Gains, Insoluble Monolayers at the Liquid-Gas Interface
(Wiley-Interscience, New York, 1966).
10. For example, see R. D. Klausner and D. E. Wolf, Biochemistry 19,
6199 (1980).
directions by a right-hand/left-hand leadscrew. The performance of this
system will be discussed and compared with that predicted by ray-tracing
calculations. The magnitudes of various lens aberrations will be com-
pared, and their effect on spatially resolved measurements of emission
from an inductively coupled plasma will be examined.
Index Headings: Optics; Lenses; Aberrations.
INTRODUCTION
Emission m o n i t o r e d b y l a b o r a t o r y i n s t r u m e n t s usually
requires an image t r a n s f e r s y s t e m consisting of mirrors,
lenses, or other optical c o m p o n e n t s . T h e r e are m a n y rea-
sons for this r e q u i r e m e n t . ~ S e p a r a t i o n of the source a n d
the detector is usually necessary because the source is
APPLIED SPECTROSCOPY

Fro. 1. Photographof the lens system.The labeled componentsare:
(1) collimatinglens, (2) focusinglens, (3) iris, (4) 4-axis lens mounts,
(5) leadscrew,and (6) dial indicatorfor focusing.
hot, at high potential, or bulky. In both routine analytical
measurements and fundamental studies of emission
sources it is often desirable to collect the radiation from
a specific region and exclude the surrounding regions.
This procedure requires that the image formed by the
transfer optics be a faithful representation of the source,
a requirement that is met only if careful attention is given
to the design and use of the image transfer system.
There is a frequent need for high-resolution optical
imaging systems in fundamental and applied studies of
sparks, arcs, inductively coupled plasmas, and other dis-
charges. 2 In fundamental studies, the spatial variations
in intensity contain information about the basic nature
of the source. In analytical measurements, the spatial
resolution is required for selection of a region in the
source which exhibits the optimum characteristics. Re-
fractive optics are appealing because of their low cost,
relative to that of mirrors, and because they can be used
in simple mounts on the optical axis. They are often used,
however, with little regard for the effect of lens aberra-
tions on image fidelity. It is our aim in this paper to
provide, with the use of examples from a lens system in
our laboratory, an indication of the capabilities and lim-
itations of refractive optics for spectroscopic image trans-
fer. This study is not intended to be a comprehensive
discussion of geometric optics. Such discussions have
long been available in a variety of sources2 -6 It is in-
tended, rather, to be an illustrative example of the use
of refractive optics in a practical, spectroscopic appli-
cation.
For imaging of small regions near the optical axis,
chromatic and spherical aberrations are of primary im-
portance. Spherical aberration is the only monochro-
matic aberration which occurs when a spherical optic is
illuminated on-axis. 1 The farther from the optical axis
that a ray enters the lens, the nearer to the lens it focuses.
There are two types of spherical aberration: longitudinal
(LSA) and transverse (TSA). The distance along the op-
tical axis between the intercept of the rays near the op-
tical axis (paraxial rays) and the rays which cross the
edge of the lens (marginal rays) is defined as LSA. TSA
occurs where paraxial and marginal rays intercept the
paraxial plane at different heights. With both LSA and
TSA, the effect is dependent upon the shape of the lens. 7
Chromatic aberration occurs when light rays pass through
a curved optic where the index of refraction is a function
of the wavelength. Instead of a well-defined image, a
series of monochromatic images come to focus, each at
a different distance along the optical axis. This paper
concentrates primarily on the effects of chromatic and
spherical aberrations. For focusing of extended sources
on an imaging detector, off-axis aberrations must also be
considered, and will be treated briefly.
EXPERIMENTAL
The lens system (Fig. 1) consists of two piano-convex
fused-silica lenses (Melles Griot, Irvine, CA) with nom-
inal focal lengths of 300 mm. The back focal length, the
distance from the focal point to the planar surface of the
lens, was reported to be 296.4 mm. With the use of the
nominal back focal length, the calculated point of best
focus for a 5.0-cm aperture is 290.4 mm. The actual back
focal lengths of the lenses were 284 mm and 282 mm.
The back focal lengths were measured by placing a plane
mirror on the curved side of the lens, perpendicular to
the optical axis. A point source was moved along the axis
on the planar side of the lens until its reflected image
was in focus at the source. The measurements were made
with a lens aperture of 5 cm. The values measured were
therefore within the _+5% tolerance for the lenses as
quoted by the manufacturer.
The lenses are mounted on translational stages that
are driven in opposite directions by a right-hand/left-
hand leadscrew. The spacing between the lenses is ad-
justed with wavelength, so that the first lens always acts
as the collimating lens and the second as a focusing lens.
The lenses are oriented so that the two convex surfaces
face each other. An adjustable iris mounted between the
lenses is used to control the amount of light permitted
to pass through the lenses.
Two emission sources were used in this study. For
characterization of the lens system, an Air Force test
target was illuminated from behind with a 4-W, low-
pressure mercury lamp. The Air Force test target consists
of parallel bars etched into an opaque chromium back-
ground on a glass substrate. To demonstrate the practical
laboratory applications, we used an inductively coupled
plasma as the source. A 1000-ppm solution of Zn was
aspirated into the plasma, and spatial profiles of the
Zn(I) 213.856-nm line were recorded. Light from both
sources was imaged on the entrance slit of a McPherson
(Acton, MA) Model 2061 monochromator. Entrance and
exit slits were set to 20 ~m. Light emerging from the
monochromator was detected with a Hamamatsu (Mid-
dlesex, NJ) R446 photomultiplier tube, and the resulting
APPLIED S P E C T R O S C O P Y 609
~10 z 1.0"
l 1 T
o1 B ./" []
D O

D
O O D
0.5 O
z .o~-:_~. A
O D
. / . . I - ~ .I.
D [] O D D D D []

O D O DD DO ~ D
D
_E
>- 13
D Cl D DD DD D D

D D t~ D D D [] 13
[] D

[]
-0.5 [] El
1:1 D

[] O

______J J_ L -1.0 , , ~ , , I I I L , , , ,
0 0 - - 2.00 4.00 S,00 8.00 -1.6 - 0.8 0 0'.8 1.6
AXIAL POSITION (mm)
Xlmm]
FIG. 2. Longitudinal ray profiles. The circles of least confusion are
FIG. 4. Transverse ray profile--f/6 at the paraxial focal point. Com-
labeled for: (A)f/7.4, (B)//11.8, and (C) f/30.
pare with Fig. 3.

photocurrent amplified by a Kiethley (Cleveland, OH)

Model 427 current amplifier. Signals were recorded with and the target with motorized stages, so that the image
a Nicolet (Madison, WI) digital oscilloscope, Model 4094. traveled across the entrance slit of the monochromator.
With the use of the ray-tracing equations from Nuss- The resultant pattern is a convolution of the image of
baum, s a computer ray-tracing program was written to the test target and the monochromator entrance slit.
generate longitudinal and transverse ray diagrams for a Lens spacing and iris diameter were changed to dem-
simple optical train. Uniform illumination of the first onstrate the effects of the lens aberrations.
surface of the optical train by a point source was simu- Because the mercury lamp produces an ac output at
lated as fol]Lows: The cone defined by the point source 120 Hz, the current amplifier was operated with a rise
and the circular edge of the optic was located, and a time of 100 ms, to filter out the 120-Hz component of
bundle of rays was generated that intersected the right the signal. This procedure required that the stages be
cross section of the cone in a uniform hexagonal pattern. moved slowly (approximately 0.16 mm/s), so that the
Although this approach is subject to spherical distortion amplifier could track the rising and falling signals as the
at large acceptance angles, it is a good approximation for alternating light and dark regions passed across the en-
the angles used in our calculations. The computer pro- trance slit.
gram provided a means of rapidly and accurately as-
sessing the effects of aberrations for different lens con- RESULTS A N D D I S C U S S I O N
figurations. Particular attention was given to the lens With the use of the ray-tracing program, a longitudinal
configuration in Fig. 1. ray profile was obtained for the region surrounding the
We assessed experimental lens performance by ad- paraxial focal point (Fig. 2). This figure represents the
justing the NIcPherson monochromator to monitor a se- effects of longitudinal spherical aberration by illustrating
lected mercury line. The Air Force test target was illu- the points of best focus for f numbers of 7.4, 11.8, and
minated with the mercury lamp creating an image of 30. The f-number is the ratio of the focal length of the
vertical bar:s. We checked image fidelity at wavelengths lens to the effective lens diameter, which is limited by
of 365.01, 435.84, and 546.07 nm by moving the lamp
0.2
1.0-

OD O~

0.1 Dee ~o~-D

~D
0.5

X 20
E 0 F , ~ I O microns~
_Eo
>-
>-

-0.1
0.5

-0.2
-0.3 "0:2 -0:1 0 0'.1 0'.2 0.3
I i J i i i I i i i i i i i
1"01.6 -0.8 0 0.8 1.6 X[mm]
Xlmml FIG. 5. Transverse spherical aberration--f/30. The inset has been
FIG. 3. Transverse ray profile--f/6 at the circle of least confusion. magnified by a factor of 20.

610 Volume 42, Number 4, 1988

 0.2 1.0-

O D o H
0.1 0.5 o
o o p
o o

O o OODO oc~ oD
O [] O o o o
O o
[] o
E E } []
>-
[] D D
D
D D [] [] [] D
O o DDDD OO13O DD
O D
- 0.1 -0.5 D [] 1:1
D

cl D D D

-0.2
-o.3 -o'.2 -o3 o d.1 0'.2 0.3 "1"01.6 ' . . .- 0 .' . 8. 0
. . . . .0.8 . . 1.6
Xlmml Xlmml
FIG. 6. Transverse spherical aberration--f/12. FIG. 8. Transverseray profile--f/6, obtained by reversing the lens
orientation so that the two planar surfaces face each other. Compare
with Fig. 3.
the adjustable iris. The plane of best focus is generally
defined by the intersection of the marginal rays with rays
arriving from a circle having a diameter of approximately and at the circle of least confusion. The f number was 6
70% of the lens diameter. 7 This plane is commonly called for both profiles. The spot sizes at the paraxial focal point
the plane of best geometric focus. The blur circle (circle and the circle of least confusion were 1.6 mm and 0.4
of least confusion) has its minimum diameter at this mm, respectively.
plane. The circles of least confusion are labeled for each Figures 5, 6, and 7 demonstrate the effect of varying
of the three f numbers. Notice that as the iris diameter the f number on the optimum spot size. Each figure shows
is increased to allow more light to pass through the lenses the intersection of the rays with the plane of best focus.
the effective focal point moves closer to the lens. The The spot diameters for the f numbers of 30, 12, and 6
paraxial focal point is located very close to the circle of are 0.0045, 0.070, and 0.37 mm, respectively. Instead of
least confusion of an f number of 30. As the f number
decreases, the marginal rays cross the optical axis farther
from the paraxial focal plane. From this figure it is also
obvious that the spot size changes as the image is de-
tected at different points along the optical axis.
Transverse spherical aberration can significantly affect
the image spot size if the image is detected at the paraxial
focal point, as opposed to the circle of least confusion.
TSA increases as the f number decreases. To demon-
strate TSA graphically, we calculated the position of the
circle of least confusion and generated a lateral profile
showing the intersection of the rays with the plane of
best focus. Figures 3 and 4 show the relative spot sizes
, , i , = ,
of the point source's image at the paraxial focal point 4 6 10

0.2.
[3 D
0 O
0 D
[] []

o
0.1 o o []
D
D o Do DO []
D
o O O O O O
o D
rj D
%
E o D
D
E~ []
D []

D []
D D
[] []
D D DD oo
tJ DD NO D
D D O
-0.1 D
o
O

D 0 J
[] D
13 o 4 6 8 10
0 [] LATERAL POSITION [ m m l
-0.2 FIG. 9. Best lateral profile resolutionusing the Air Force test target
-o.3 -0:2 -0:1
0 0'.1 0'.2 0.3
at the 435.8-nm mercury line: (A) f/30, (B) f/8.7. The bar widths for
Xlmml the six groups from left to right are: 0.140, 0.158, 0.177, 0.198, 0.223,
FIG. 7. Transverse spherical aberration--f/6. and 0.250 mm.

APPLIED SPECTROSCOPY 611


= 0
4 6 8 10
LATERAL POSITION [mm]
FIG. 10. Lateralprofiles of the Air Force test target at 546.1 nm: (A)
best experimentalfocus, (B) white light focus.
perfect point images, laterally blurred images are formed.
At f/6, two points in a source must be separated by 0.37
mm to be completely resolved.
In addition to aperture, lens shape and orientation play
important roles in spherical aberration. Generally, sim-
ple positive lenses have under-corrected spherical ab-
erration; negative lenses are usually over-corrected. 7 Op-
timum correction is achieved by using more than one
optical element. For a single-lens system, the best shape
for 1:1 imaging is a symmetric biconvex lens. A dramatic
improvement in performance over single lens systems is
gained by using two piano-convex lenses with the convex
surfaces facing each other. Figure 8 shows the effects of
reversing the lenses from their optimum orientation. The
improvement in spot size in the two-lens system, com-
pared with a single-lens system, is a factor of 3.6.
We demonstrated the experimental effects of spherical
aberration by first adjusting the monochromator to the
most intense mercury line (435.84 nm). The lenses were
adjusted so that they were spaced according to the cal-
culated po~,~itionof best focus. The test target was placed
between the lamp and the lenses, and lateral profiles were
taken for iris diameters of 10.0 mm and 50.0 mm. Figure
9 illustrates the best resolution for each of these f num-
bers (8.7 and 30). The values of 8.7 and 30 are the prac-
tical limits of the instrument. The monochromator is an
]/8.7 instrument, and above [/30 the throughput is too
low for practical measurements. At ]:/30 the bars in the
test target are nearly baseline resolved and the peak
intensity is uniform across the width of the scan. The
peaks approach the ideal trapezoidal shape that would
be predicted from the convolution of the undistorted bar
612 Volume 42, Number 4, 1988
A
o ~ 10
O 4 6 8 10
LATERAL POSITION {mml
Fro. 11. Lateralprofilesof the Air Force test target at 365.0 nm: (A)
best experimentalfocus, (B) white light focus.
pattern with the entrance slit. At [/8.7 the effects of
spherical aberration are noticeable, particularly for the
smaller bar spacings. The peaks are no longer completely
resolved and have a triangular shape, rather than the
ideal trapezoidal shape.
The effects of chromatic aberration are usually more
dramatic than those of spherical aberration. In atomic
spectroscopy, most measurements are made in the wave-
length range between 200 and 800 nm. Inexpensive ach-
romatic lenses are available from several sources (e.g.,
Newport Corp., Fountain Valley, CA) for the visible light
region from 400 to 800 nm. To cover the ultraviolet region
(200-400 nm), one must employ special UV achromats.
The selection of such lenses is limited, and their cost can
be prohibitive. The only option for coverage of the entire
spectral region is an adjustable system composed of sin-
gle-element lenses.
Figures 10 and 11 demonstrate the results of chromatic
aberration with the use of the AF test target and the
mercury lamp. There is a noticeable difference between
the profiles taken at the calculated best focus and the
white light focus. We obtained the white light setting by
illuminating the AF test target with a tungsten lamp and
adjusting the lenses by hand until the best focus by eye
was achieved on the entrance slit of the monochromator.
This setting corresponded to a wavelength of 616.0 nm.
For the 546.1-nm mercury line the effect is very similar
to the spherical aberration effects discussed earlier in the
paper. The effects for the 365.0-nm line are even more
dramatic. Not only are the bars within each group blurred
together, but the groups themselves are not resolved.
Compensation for chromatic aberration is essential for
 -19.7

[]
>- 113D
i- lO -19.8 ~3OoI O [3
U) O []
Z
U.I OOo E
I-- IoOoOD
_z E O OODE ID O
W
> _E
>-
-19.s DoOo[ IDODD D
OoDoE IDoDO D
OoOoO IIDDDo O
i,i
n- Ol3O[]lIDDOD
-20.0 [:30[31IDDD

O i i i -20.I
o.s o:2 o'.1 o o:1 o'.2 o.s
LATERAL POSITION (m m) Xlmml
Fro. 12. L a t e r a ] profi]es o f t h e Z n ( I ) 213.856-nm emission line at 4 FIG. 13. Twenty millimeter (3.9 degrees) off-axis transverse profile--
m m above t h e ]oad coil: (A) calculated best focus, (B) white light focus. f/30. Compare with Fig. 5.

resolution of even the coarsest features of a source, par-
ticularly if it is emitting in the ultraviolet.
To illustrate this point with a commonly used source,
we used our lens system to record the spatial profile of
the Zn(I) emission line from the ICP. Figure 12 illustrates
the images obtained from the zinc line at the calculated
focus and a white light focus. The effects of chromatic
aberration were, as predicted, dramatic. The intensity of
the correctly focused profile was more than twice that of
the profile focused by eye. The intensity difference is
marginally important; the major effect is the loss of spa-
tial definition in the profile. The correctly focused profile
shows cooling in the center of the plasma, which is known
to exist. The white light focused profile is void of almost
all spatial features.
Astigmatism and coma are aberrations which arise when
an optic is illuminated off-axis. Our system is designed
completely on-axis. However, for instruments such as
imaging spectrometers, which use the entire image, off-
axis aberrations must be considered, and we include a
brief treatment of them here.
Coma begins to occur when an optic is illuminated at
small off-axis angles. It is due to the property that the
lateral magnification of the image is dependent upon the
distance from the optical axis at which the incoming rays
strike the optic? It is similar to spherical aberration, in
that the rays striking the optic farther from the optical
axis come to a focus closer to the lenses than do the rays
passing through the lens nearer to the center. The image
formed from a point source illuminated off-axis is diffuse,
being made up of superimposed "comatic circles" that
isotropically from a point source 20 mm off-axis (3.9
degrees). In Fig. 13 the aperture was limited to 10 mm
in diameter. At this small aperture, the predominant
aberration shown is astigmatism. The rays cross through
the focal plane in an elliptical shape. Even more impor-
tant, the spot size is many times the size of the same
profile taken on-axis. For Fig. 14 the aperture was set to
50 mm, increasing the contribution of coma. Close ex-
amination of the profile reveals the series of comatic
circles mentioned previously. As is the case with the
previous figure, the spot size is many times larger than
the same profile taken on-axis. Off-axis aberrations can
significantly affect image transfer for applications using
lenses, even at small off-axis angles. Their effects must
be considered in the design of an imaging spectrometer
that uses refractive optics.
CONCLUSIONS
If proper attention is given to the elimination of lens
aberrations, submillimeter spatial resolution can easily
be achieved in refractive imaging systems used close to
the optical axis. For extended sources (such as the ICP)
that lack sharp boundaries, adequate resolution can be
-18.5-
D D
[] D
-19.C D
[] D o

o [3
combine to give it a comet-like appearance.
The effects of astigmatism increase nonlinearly with D D • D D D

increasing displacement of the source from the optical

axis. From a point source, astigmatism results in a pair
of sharp, perpendicular lines being formed at separate D D [] []
axial positions, instead of a perfect point image. In be-
-20.C D ~ D
tween these two perpendicular lines the image formed
[] []
appears as either an elliptical or a circular blur. The
amount of astigmatism is the distance between the two
conjugate lines. Astigmatism and coma depend strongly -1.6 . . . . - 0.8
. . . . . . 0 o'.8' ' ' 1.6
on the conjugate ratio at which the lens is being used. 7 Xlmml
Figures 13 and 14 demonstrate the effects of these off- FIG. 14. Twenty millimeter (3.9 degrees) off-axis transverse profile--
axis aberrations. In both cases the rays were generated f/6. Compare with Fig. 3.

APPLIED SPECTROSCOPY 613

obtained by the elimination of chromatic aberration. For
compact sources such as microwave plasmas or sparks,
adequate resolution can only be obtained by the elimi-
nation of chromatic aberration and the careful minimi-
zation of spherical aberration. This minimization in-
cludes the correct choice of lens form and orientation,
the use of small apertures, and the a d j u s t m e n t of focal
point, with changes in aperture to compensate for trans-
verse spherical aberration.
T h e lens system described in this paper was designed
for use in p a i n t - t o - p o i n t imaging on the optical axis. If
a lens system is to be used for high-fidelity imaging of
an e x t e n d e d source on an imaging detector, the off-axis
aberrations of coma and astigmatism must also be con-
sidered. Like spherical aberration, coma can be mini-
mized by the proper choice of lens configuration and the
use of small apertures. T h e same is not true of astig-
matism. T h e user of a simple lens system must either
The Combination of High-Resolution Molecular Fluorescence
Spectroscopy with Liquid Chromatography and
Thin-I,ayer Chromatography*
J . W . H O F S T R A A T , t C. G O O I J E R ,
Department of General and Analytical Chemistry, Free University, De Boelelaan 1083, 1081 HV Amsterdam, the Netherlands
Under certain conditions, highly specific fluorescence spectra, showing
molecular vibrations as in infrared spectroscopy, can be obtained for the
analysis of compounds separated in TLC and microbore LC. Two tech-
niques, both employing low-temperature solid samples, are available:
Shpol'skii spectroscopy and Fluorescence Line-Narrowing (FLN) spec-
troscopy. The former utilizes n-alkane solvents, which form regular,
crystalline solids; the latter is based on selective laser excitation. Both
techniques can be applied for detection in TLC. FLN is more feasible,
since the Shporskii method requires spraying of the TLC plate, which
leads to significant spreading of the spots. FLN can also be used for off-
line detection in micro-LC after immobilization of the column effluent
on a TLC plate. In addition, some exploratory experiments on the ap-
plicability of FLN for on-column detection in LC are presented.
Index Headings: Fluorescence line-narrowing spectroscopy; Shpol'skii
spectroscopy; Thin-layer chromatography; Liquid chromatography.
INTRODUCTION
F l u o r i m e t r y is a widely applied m e t h o d for analysis of
organic compounds. It is obviously not as universally
applicable as UV/Vis absorption spectroscopy b u t is, in
principle, m u c h more sensitive. F u r t h e r m o r e , since two
Received 3 August 1987; revision received 2 November 1987.
* Presented at the XIIIth Annual Meeting of the FACSS, St. Louis,
Missouri, 1986.
t Author to w:hom correspondence should be sent. Present address:
Ministry of Traffic and Public Works, Department of Public Works,
Tidal Wateri~ Division, Nijverheidsstraat 2, 2288 BB Rijswijk, the
Netherlands.
614 Volume 42, Number 4, 1988 0003-7028/S8/4204-061452.00/0
© 1988 Society for Applied Spectroscopy
work with the off-axis image degradation caused by as-
tigmatism or consider alternative image transfer systems.
ACKNOWLEDGMENT
This material is based upon work supported in part by the National
Science Foundation under Grant No. CHE-8501555.
1. S.A. Goldstein and J. P. Walters, Spectrochim. Acta 31B, 201 (1976).
2. D. M. Coleman and M. A. Sainz, Appl. Spectrosc. 35, 125 (1981).
3. C. S. Williams and 0. A. Becklund, Optics: A Short Course for
Engineers and Scientists (Wiley-Interscience, New York, 1972).
4. M. V. Klein and T. E. Furtak, Optics (John Wiley and Sons, New
York, 1986), 2nd ed.
5. W. T. Welford, Optics (Oxford University Press, New York, 1981),
2nd ed.
6. W. T. Welford, Aberrations of Optical Systems (Adam Hilger Ltd.,
Bristol, Great Britain, 1986).
7. Melles Griot, Optical Guide 3 (Melles Griot, Irvine, California, 1986),
p. 17.
8. A. Nussbaum, Am. J. Phys. 47(4), 351 (1979).
and N . H . V E L T H O R S T
wavelengths can be chosen for the d e t e r m i n a t i o n of a
c o m p o u n d (i.e., one for excitation and one for emission),
fluorimetry is also more selective. However, the tech-
nique is almost exclusively used in quantitative analysis.
In general, the fluorescence spectra are not sufficiently
specific for identification purposes: in fluid samples and,
for the most part, in solid solutions, and for larger mol-
ecules in the gas phase as well, t h e y are broad b a n d e d
and relatively featureless.
T h e reasons for the lack of structure in such molecular
spectra differ, depending upon the state of aggregation
of the sample. In all cases the minimally achievable band-
width is d e t e r m i n e d by the lifetime of the states involved
in the transition. This ultimate, so-called natural, band-
width is given by the Heisenberg u n c e r t a i n t y relation,
according to which the u n c e r t a i n t y in energy of a state
with lifetime r is given by A E = h/2~rr J, where h is the
P l a n c k constant. For fluorescence transitions the lifetime
of the excited state usually is in the low nanosecond
range, leading to a natural b a n d w i d t h in the order of
10 =5 nm. In practice, such b a n d w i d t h s are rarely mea-
sured. In the gas phase, spectral bands are m u c h wider,
as a consequence of Doppler broadening and vibrational
and rotational congestion effects evoked by the relatively
high t e m p e r a t u r e s generally applied. In liquid solutions,
the broadening is mainly caused by the rapidly varying
interactions of dissolved molecules with the solvent mol-
ecules surrounding them, which lead to shifts in the sol-
APPLIED SPECTROSCOPY