Textile Research Journal Article

Automatic Volumetric Measurement of Nanofiber Webs using

Metaball Approximation Based on Scanning Electron
Microscope Images
In Hwan Sul
Abstract Scanning electron microscope (SEM) i-Fashion Technology Center, Konkuk University, Seoul,
images are an essential tool for identifying the 143-701, Republic of Korea
microstructures of fibrous and polymeric materi-
als. Useful information, including fiber orienta- Young Jun Cho
tion and radius distributions, can be retrieved Hyosong R&D Business Labs., Gyeonggi-do, 431-080,
from the SEM images. However, the image is two Republic of Korea
dimensional and its manual measurement is inevi-
table for acquiring volumetric information. This
Chang Kyu Park1
paper reconstructed the three-dimensional shape of
Department of Textiles and Engineering, College of Eng.,
the fibers by approximating the fibers with spheres
Konkuk University, 143-701, Republic of Korea
known as metaballs. Image analysis techniques were
used to generate metaballs and the volumetric
information was calculated from the sizes and posi-
tions of the metaballs. The proposed method was
tested with nano-scale polymer SEM images and
automatic statistical measurements were compared
with the manual measurements.

Key words scanning electron microscope image,

metaball approximation, orientation vector, radius
distribution, pore size distribution

The scanning electron microscope (SEM) is a device that
uses high energy electrons to visualize the material surface
topography. It has been used in wide areas, ranging from
biology to the material sciences. In particular for textiles
and polymeric materials, we can perform both qualitative
and quantitative observations on a specimen by taking
SEM images. For qualitative measurements, the SEM can
produce the surface status, such as smoothness, and
researchers can judge if the synthesis was successful or not.
For quantitative measurements, we can measure volumet-
ric sizes, such as fiber orientations, fiber radius distribu-
tion, and pore size distribution.
Meanwhile, most specimens used for SEMs have a
three-dimensional microstructure, such as polymers.
Since a SEM image can give only a two-dimensional pic-
ture, researchers should measure fiber sizes manually and
predict their true values using statistical methods, such as
stereology [1]. A stereology-based method is useful for
retrieving the size information, such as the volume fraction
from two-dimensional cross-section information, e.g. for
finding the volume fraction of particles in reinforced mate-
rials. However, the stereology-based method is classified as
an indirect method because it is based on statistical infer-
ence. Moreover, the manual measurement of fiber radii is
a laborious job. This paper proposes an automatic method
of acquiring statistical values from a single SEM image by
reconstructuring the original micro-fibers in three dimen-
sions. We assume that the specimens have three-dimen-
sional web-like fibrous structures of multiple overlapped
layers of nano-scaled fibers. Hereafter, these fibers are
referred to sometimes as fibers, micro-fibers or nanofibers,
1
Corresponding author: tel: +8224504196; e-mail: cezar@
konkuk.ac.kr

Textile Research Journal Vol 80(11): 995–1003 DOI: 10.1177/0040517509348333 © The Author(s), 2010. Reprints and permissions:
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TRJ 996 Textile Research Journal 80(11)
and their SEM images are referred to as fiber SEM
images.
Reconstructing the three-dimensional shape of the
material microstructure has been an interesting theme for
many material scientists. The well-known technique is a
serial sectioning-based method using a focused ion-beam
scanning electron microscope (FIB-SEM). Several hun-
dred tomographic SEM image slices with a thickness of a
few nanometers can be acquired through a focused ion
beam (FIB) sectioning process [2]. Next, a three-dimen-
sional shape can be reconstructed by stacking the images.
Sidhu and Chawla reconstructed the three-dimensional
image of Ag3Sn by serial section [3] and measured the
aspect ratio. Li et al. [4,5] also quantitatively characterized
the microstructural damage of Si particle-reinforced metal
matrix composites by using serial sectioning. They calcu-
lated a particle size, a shape, an orientation, a local volume
fraction, and neighbor distances from the three-dimen-
sional reconstructed data. The reconstructed three-dimen-
sional data were also useful as mesh data for further
analysis using finite element analysis [6].
The specimens for the slicing methods had only a single
object, such as particles, rods, or chromosome [7], so each
sliced image can be merged and reconstructed relatively
easily. However, the same technique cannot be simply
adapted to the SEM images of the polymeric materials.
The major problem is that the polymers have a complex
structure with multiple layers of interconnected fibers, so
that the sliced images still contain multiple objects that
cannot be easily isolated.
There were also other approaches that used several
pieces of the SEM images without slicing them. Samak et
al. [8] used two pieces of the SEM images and visualized
the surface ruggedness of the ceramic and the synthetic
foams with a stereo-matching algorithm. They increased
the visual quality by means of texture mapping, but they
did not deal with the quantitative measurement. Palus-
zyński and Slówko [9] also used four detectors for rela-
tively smooth objects and it seemed to be difficult to apply
to a complex polymeric structure. Those methods used a
single object specimen and were not concerned with the
isolation or the qualitative measurements.
The SEM is an innovative device for showing the micro-
structure of materials. However, mechanical engineers
have no proper tools to analyze the captured SEM image,
except for using a ruler. A fiber SEM image is composed of
hundreds of fiber segments and each fiber has a different
radius. Even a single fiber has different radii depending on
its shape. Thus, it is not practical to use only the manual
measurement for acquiring a statistically true average
value. This paper presents a novel way to acquire a large
amount of measurement data samples from a SEM image by
separating the overlapped fiber images and reconstructing
the three-dimensional mesh. This paper adopted several
image analysis techniques to partition the two-dimensional
SEM image into groups of fibers and used metaball-based
approximation to build three-dimensional mesh from the
two-dimensional fiber images. Once the metaballs’ posi-
tion and radii are acquired, the statistical values can be cal-
culated easily. In addition, there were some limitations in
our approach. Only the top-most layer of fibers could be
easily identified, although general fiber SEM images are
composed of multiple numbers of layers. A high-resolution
SEM image was preferred because it contains less noise. To
simplify the algorithm, we assumed the cross-sections of
fibers to be circular. Our algorithms are described in detail
in the following.
Algorithm
Two-dimensional Image Analysis Procedure
The basic idea of our method with respect to a simple
object is illustrated in Figure 1. Suppose that the check-
mark-like letter of Figure 1(a) is a kind of fiber and we
intend to measure its dimensions. We want to find simple
geometric features, such as a series of circles, to represent
the checkmark. To do so, the center line of the checkmark
is acquired first (Figure 1(b)) and the circles are deployed
along the center line (Figure 1(c)). Lastly, the circles are
converted to three-dimensional spheres so that the finally
reconstructed mesh can be produced (Figure 1(d)). How-
ever, since the fiber SEM image has many overlapped
lines, further treatments are needed in fact.
In the beginning, we need several assumptions to sim-
plify the problems. The first assumption of our method is
that the pixel brightness is proportional to the actual z-
coordinate of the specimen. This can be readily proven
from the mechanism of the SEM. The first-hand objective
of finding z-coordinates from pixel values is to distinguish
each fiber and get the true radii, not to make the final
three-dimensional mesh. Fiber SEM images are composed
of hundreds of inter-connected fiber segments. Without
separating each fiber, the radii have the wrong values, as
Figure 2(e) shows. The closer (higher z-coordinate) object
to the SEM detector has a brighter pixel (high binary value
close to 255) and vice versa. Therefore, fibers closest to the
detector will have white colors in the SEM image. Our idea
is to collect pixels of similar colors and identify them as a
group of fibers preliminarily. As our objective is the quan-
titative measurement, such as the radius distribution, this
grouping does not have to be as perfect as a human eye
recognizes the fibers. The grouping has only to disassemble
the stacked fiber images into individual single layers to
ease the whole procedure. To this end, a series of image
analysis methods were applied to the original SEM images.
The second assumption is that we use only the top-most
layers of the fibers. This is an evident shortcoming of our
method, but there seems to be no easy way to completely
 Automatic Volumetric Measurement of Nanofiber Webs using Metaball Approximation Based I. H. Sul et al.
Figure 1 Diagram of the reconstruction algorithm: (a) origi-
nal two-dimensional picture of a three-dimensional object;
(b) center line (skeletonization); (c) metaball approximation;
(d) three-dimensional mesh generation.
identify the fiber layers with image analysis techniques.
Nonetheless, we believe that our method is more effective
than the manual measurement.
In this two-dimensional image analysis procedure, the
first objective is to distinguish fibers into several groups.
Figure 2(a) shows the original SEM image. As is the usual
case in the fiber webs, the several layers of the fibers are
overlapped and it seems difficult to distinguish them with
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Figure 2 Image analysis procedure for sample #PUM1 ((a)
original image; (b) histogram equalized; (c) smoothed; (d)
thresholded; (e) skeletonized; (f) Canny edge; (g) flood filled;
(h) skeletonized from (g); (i) final metaballs).
conventional methods. As a first step, the image was equal-
ized (Figure 2(b)), smoothed (Figure 2(c)), and thresh-
olded (Figure 2(d)) to convert the gray image into a binary
value of 0 or 255. Histogram equalization was used to
enhance the image and to make the inter-fiber interface
clearer. The raw image had similar gray values, but the
equalized image exhibited the gradient difference among
the fibers. The thresholded binary image (Figure 2(d)) was
used to get the center lines (skeletons) of the fibers using a
thinning algorithm by Lee et al. [10], as in Figure 2(e).
However, the fibers generally had scales on their surface
and these scales acted as unnecessary noise groups. So, the
images needed to be smoothed (Figure 2(c)) before being
thresholded. Gaussian smoothing with a radius of 3 pixels
and a threshold value of 128 was used. However, as seen in
Figure 2(d), a simple binary image after thresholding does
not distinguish the overlapped fibers, so the skeletons of
Figure 2(e) can be said to be a wrong result, especially in
the regions where fibers are overlapped. The cause is that
we tried to treat the bulk image all at once.
Thus, we rejected Figure 2(e) and skeletonized one
fiber group at each time instead. Herein, we define a col-
TRJ 998 Textile Research Journal 80(11)
lection of pixels with similar colors in the same fiber seg-
ment as a fiber group. To dissect the original binary bulk
image to multiple fiber groups, the raw image was edge
detected with the Canny edge algorithm [11] from the histo-
gram-equalized gray image (Figure 2(b)). Figure 2(f) shows
the edge detection result, which preserves the inter-fiber
boundary well. Figure 2(f) depicts a collection of simply-con-
nected closed-loop polygons. Then, each fiber was painted
with different colors with a flood-fill algorithm [12]. The out-
of-fiber regions (pore or background regions) were not
painted because they were not necessary. Determination of
the in/out of the fibers was based on the binary color of Fig-
ure 2(d). Figure 2(g) shows the distinguished fiber groups.
Then, the skeleton lines of each group were detected (Fig-
ure 2(h)). In Figure 2(e) the skeleton lines were not in the
exact centers of the fiber groups. In contrast, those of Fig-
ure 2(g) resided in the correct middle lines. As our grouping
is based on the pixel colors, there are some errors in the
grouping, i.e. a single fiber is broken into several groups, not
into a single group. However, this does not make any dif-
ference because the metaballs are finally converted into
the same mesh in the mesh generation procedure using the
marching cube method.
Three-dimensional Metaball Approximation
Once the overlapped fibers were distinguished, the rest of
the job was to approximate the fibers with simple geometry,
such as a sphere. As we have only a single SEM image as our
input data, we cannot make three-dimensional meshes
directly as in the FIB-SEM serial sectioning method. Here
we define another assumption – that the fibers have the
same cross-sectional shape. Generally, the polymeric or
fibrous material has circular or elliptical cross-sections. In
fact, any kind of cross-section shape can be used, but we
chose the simplest one to ease the calculation. Once the
cross-sections are the same, the individual fibers can be
modeled with the same basic geometric feature, such as
cylinders or spheres of various sizes and directions. We
approximated the fibers as a bunch of overlapped balls
with different radii, as seen in Figure 2(i). These balls are
also known as metaballs [13]. The metaball is an implicit sur-
face used to model a three-dimensional shape effectively. To
determine the positions and radii of the metaballs, the skele-
tons of the fibers were needed. The metaballs were assigned
to each pixel of the skeleton lines of Figure 2(h). The radius
of each metaball was obtained from the biggest incircle
contacting with the fiber boundary. As too many metaballs
can slow down the mesh generation process, balls within a
certain range of threshold distance were omitted. Figure
2(i) is the two-dimensional metaball insertion result. They
were given x and y coordinates but not z-coordinates as yet.
The z-coordinate value was calculated from the sum of the
pixel values within each metaball. Thus, the metaballs in a
brighter region take higher z-coordinates and vice versa. In
simple mathematical terms, the total volume V of all the
fibers can be shown as:
f–1
V = ∑V i (1)
i=0
where f is the number of fibers.
The volume of each fiber, Vi, is:
Vi = Bi,0 傼 Bi,1 傼 … Bi,b–1 (2)
where b is the number of balls in fiber i.
The metaball Ri,j is expressed as an equation of a circle
as follows:
2 2 2 2
( x – X i, j ) + ( y – Y i, j ) + ( z – Z i, j ) – R i, j = 0 (3)
where (Xi,j, Yi,j, Zi,j) and Ri,j are the center and the radius of
metaball i,j, respectively. Each pixel of skeleton lines
becomes the metaball center and the maximum incircle
radii become the metaball radii when they touch the fiber
boundary curves. The binary image after the thresholding
was used to find the incircles. Note that the metaballs can
have different radii depending on their positions, while
they retain the same circular, cross-sectional shape. More-
over, the path of the metaballs can be curvilinear if the
original fiber group has a curvilinear shape.
Now that the positions and the sizes of the metaballs
have been found, the quantitative analysis can be done. The
radius distribution can be plotted directly from Ri,j. The ori-
entation vector O i, j of a metaball Bi,j was defined as the dis-
placement vector between two neighboring metaballs, i.e.,
O i, j = { ( X i , j + 1 – X i, j ) , ( Y i, j + 1 – Y i, j ) , ( Z i, j + 1 – Z i, j ) }
(4)
(0 ≤ j < b)
As there can be noises in the images and the metaballs with
the very small radii can affect the statistics, all the statisti-
cal values were weight-averaged with the size of the meta-
ball. For example, an average orientation angle θi of a fiber
Fi was calculated as:
b–1
∑
i=0
R i, j • θ i, j
θi = ------------------------------ (5)
b–1
∑
i=0
R i, j
where θi,j is the angle between O i, j and the +x-axis.
To show the three-dimensional reconstruction result of
the SEM image, the metaballs should be converted to
 Automatic Volumetric Measurement of Nanofiber Webs using Metaball Approximation Based I. H. Sul et al.
Figure 3 Three-dimensional reconstruction result for
sample #PUM1: (a) top view; (b) front view.
three-dimensional mesh data. Marching cube [14] is the
famous technique to convert the metaball space to triangu-
lar mesh data. To use the marching cube method, the space
should be voxelized first because the marching cube
method needs voxel data as its input. So the whole three-
dimensional space containing the volume V was voxelized.
If a voxel lies inside any metaball, it is assigned a value of 1
and otherwise 0. The voxels were also used for the calcula-
tion of the fiber volume fraction. The total volume fraction
can be found simply by dividing the number of filled voxels
by the total voxel numbers. Figures 3(a) and 3(b) show the
top view and the front view of the final three-dimensional
mesh after the reconstruction.
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Table 1 Specification of the specimens used.
Item PUM1 PUM2
Material PU PU
Thickness (μm) 8–10 12–15
Mean flow pore diameter (μm) 0.6227 0.5805
Bubble point pore diameter (μm) 0.8192 0.7444
Results and Discussion
Test Specimens
To verify the presented method for the actual fiber SEM
images, two kinds of poly-urethane based breathable
nanofiber membrane samples were used. Table 1 shows
the specification of the specimens. The specimens are
made of the same polymer resins, but different electrospin-
ning conditions resulted in different orientations and
radius distributions. Firstly, we need to determine the
proper size of the input SEM image size. Too large a pic-
ture size can delay the calculation time and the possibility
that the noises are accepted as valid metaballs can increase.
By resizing the picture into smaller sizes, the noises were
smoothed and the final three-dimensional reconstruction
mesh contained only the valid fibers. Images were resized to
512 pixels × 512 pixels in this case. The SEM resolution was
the more critical factor for metaball approximation. Figure 4
shows the metaball approximation results in three different
SEM resolutions. The lower the resolution is, the more jag-
ged the skeletonized center lines become, and the final
metaballs yield the more discontinuous lines. In conclu-
sion, the sufficiently high resolution of the SEM images
was advantageous in that the fiber diameter should be
greater than a tenth of the image width. We used images of
resolution 2 × 106 times where 1 μm equals 84 pixels.
Figure 2 shows the image processing steps for the speci-
men #PUM1 and Figure 3 shows the final three-dimen-
sional mesh reconstructed as explained earlier. The top-
most layer of the original image was successfully recon-
structed. As we have the three-dimensional mesh and the
metaball information, the fiber statistics, such as radius
distribution and orientation, can be simply acquired. Some
noises exist in Figure 3, but they do not adversely affect the
statistics because the statistics are factored by the radius
size. An AMD 3.0GHz dual-core personal computer was
used and the processing time was about 30 seconds for the
512 × 512 image.
Using the manual measurement, the only information
we could infer was that the radius size would be about
75~135 nm and the average radius would be about 100 nm.
Figures 5(a) and (b) depict the fiber statistics. To get the
same graph with the manual measurement, hundreds of
TRJ 1000 Textile Research Journal 80(11)
iterations would be necessary. As more fibers participated
in the calculation compared to the manual measurement,
the resultant statistical data must be closer to the true val-
ues.
It is apparent that the volume fraction can also be calcu-
lated, because the total volume and the fiber volume are
known from the voxel data. However, we were not able to
find a material whose exact microscopic volume fraction
was known. Instead, the mean flow pore size was chosen to
verify the results. The pore size distribution of the
nanofiber membrane was determined using a capillary flow
porometer (Porous Materials Inc., Model CFP-1200AE).
As one can see in Table 1, the experimental value of the
mean flow pore diameter was 0.6227 μm and this is similar
to the result in Figure 5(c) with about 6% error. The pore
size was also easily calculated by inverting the image of
Figure 2(d) and making metaballs for pores. The theoreti-
cal pore diameter was chosen as the maximal radius of the
pore regions. The result shows that the graph from the pro-
posed method may be said to coincide with the expected
value.
The same procedure was applied to specimen #PUM2
and the results are shown in Figures 6–8. The manual
measurement values were similar to those of #PUM1. The
final three-dimensional mesh was also found and it was
Figure 4 Results for various SEM
resolutions (from left: original
image; fiber groups; metaballs;
three-dimensional mesh): (a)
×20,000 (lower resolution); (b)
×200,000; (c) ×2,000,000 (higher
resolution).
revealed that the radius distribution did not vary, but the ori-
entation graphs showed a different result from Figure 5(b).
Such a slight difference cannot be easily identified through
a few repetitions of the manual measurement. The advan-
tage of the automatic measurement is clearly shown here, in
that the more general statistical values can be known com-
pared to the manual measurement, although our method
was confined to the top surface of the specimens. The pore
size error with the experimental value was about 12% in
this case, under the assumption that the capillary flow
porometer gave a true value of porosity. We postulate that
the error was caused by using only the surface fiber infor-
mation. The error was a little bigger than that in the
#PUM1, but we succeeded in acquiring a detailed distribu-
tion of the pore size in a two-dimensional graph form,
while the porometer gives only the mean and standard
deviation values.
The shortcoming of using the high resolution SEM is
that the statistics represent only a small range of the area
from the specimen. To get the more general statistics, there
are two alternatives. One approach is to carry out the
above-mentioned procedure repeatedly over different
regions. This does not require a special technique but
needs only additional SEM filming. The other approach is
to use low-resolution images. It was not possible to gener-
 Automatic Volumetric Measurement of Nanofiber Webs using Metaball Approximation Based I. H. Sul et al.
Figure 5 Radius distribution and fiber orientation graphs
for sample #PUM1: (a) radius distribution; (b) fiber orien-
tation; (c) pore diameter distribution (mean = 587.69 nm,
st.dev. = 15.9 nm).
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Figure 6 Example of three-dimensional reconstruction
from two-dimensional SEM image for sample #PUM2.
ate valid metaballs at the low resolution as shown in Fig-
ures 4(a) and (b) because the fiber segments were too thin,
but the center lines could be found if a sufficiently larger
size of the input image was used (Figure 9). Center lines
cannot give the radius distribution, but they can offer
knowledge of fiber orientation instead. Combining the
radius distribution result acquired from the high-resolution
image with the fiber orientation acquired from the low-res-
olution image can give more general statistical values for
the given specimens. The main concern of this paper was
to replace the manual measurement with a simple auto-
matic one using a single SEM image, so the verification of
the alternative methods was skipped.
Conclusions
This paper presents a new methodology based on the
image analysis for measuring fiber statistics from a single
SEM image. The main contribution is that researchers can
have more detailed and general distribution information of
the fiber radius, the orientation, and the pore size com-
pared with the manual measurement. Unlike the FIB-
TRJ 1002 Textile Research Journal 80(11)

Figure 7 Three-dimensional reconstruction result for

sample #PUM2.

SEM-based method, our method needs only one SEM pic-

ture and the complex fiber webs are decomposed into mul-
tiple fiber groups. The original two-dimensional image is
approximated with sphere-like metaballs after the image
analysis steps. Three-dimensional voxels and mesh are con-
structed from the metaballs for visual representation and
also fiber statistical data are acquired from the metaball
information. The fiber orientation, the fiber radius distri-
butions, and the pore diameter distributions are directly
known from the metaballs and the voxels. Our method is
simple, fast, automatic, and more advantageous than the
manual measurement using the stereology because it can Figure 8 Radius distribution and fiber orientation graphs
also give fiber orientation results. It is also advantageous to for sample #PUM2: (a) radius distribution; (b) fiber orien-
the FIB-SEM method in that the individual fiber is identi- tation; (c) pore diameter distribution (mean = 521.1 nm,
fied, which is an essential step for calculating the statistical st.dev. = 11.1nm).
information.
 Automatic Volumetric Measurement of Nanofiber Webs using Metaball Approximation Based I. H. Sul et al.
Figure 9 Skeletonized center lines of sample #PUM2
(×3000 SEM, 1024 pixels × 1024 pixels).
However, there were also limitations. Low-resolution
SEM images were not applicable because the overlapped
fibers could not be easily segmented. It was only possible to
get their skeletonized center lines by using a larger image
file, which required more processing time and might con-
tain a greater number of noises. We leave the case of the
low-resolution image as a further work.
Acknowledgements
This research is financially supported by the Ministry of
Knowledge Economy (MKE) and Korea Institute for
Advancement in Technology (KIAT) through the Work-
force Development Program in Strategic Technology.
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