Anatomy & Physiology
Scanning Electron Microscopic Features of the External and
Internal Surfaces of Normal Adult Lacrimal Drainage System
Mohammad Javed Ali, F.R.C.S.*, Farhana Baig, M.D.†, Mekala Lakshman, PH.D.‡, and Milind N. Naik, M.D.*
*Dacryology Service, L.V. Prasad Eye Institute, Hyderabad; †Department of Pathology, Global Hospital, Hyderabad;
and ‡Ruska Labs, Hyderabad, India
Aim: The aim of this study was to examine the ultrastructural
features of the external and internal surfaces of healthy lacrimal
drainage systems.
Methods: A prospective interventional study was performed
on the healthy adult lacrimal drainage systems obtained from
fresh exenterated specimens. Exenteration was performed for
malignancies unrelated to lacrimal system where preoperative
lacrimal evaluation was normal. A careful and thorough
dissection was carried out to isolate the entire lacrimal drainage
system from the punctum to the nasolacrimal duct. The analysis
was performed using the standard protocols of scanning electron
microscopy.
Results: Inner punctal surfaces showed a definite and slightly
elevated junction between the luminal surfaces of punctum and
beginning of the vertical canaliculus. Similar junction could
be identified between the lacrimal sac and nasolacrimal duct.
The valves of the canaliculi showed broad rugae-like mucosal
surfaces, whereas the external surfaces of the canaliculi
demonstrated well-defined orbicularis muscle with collagenous
attachments. The walls of the lacrimal sac and nasolacrimal duct
showed dense vascular plexus which included wide luminal
arteries, throttle veins, and large capacitance vessels.
Conclusions: Ultrastructural features of external and internal
surfaces of lacrimal drainage system help in better understanding
of its anatomy and physiology. The junctional area between the
punctum-vertical canaliculus and lacrimal sac–nasolacrimal
duct needs further exploration to understand their roles.
(Ophthal Plast Reconstr Surg 2015;31:414–417)
U ltrastructural features of lacrimal system and its disorders
is not very well studied.1–10 Most of the studies used com-
binations of scanning and transmission electron microscopic
techniques, however very few studied the scanning electron
microscopic (SEM) features of normal systems.3,6,9,10 The earli-
est detailed study was by Radnot in 19721 followed by Adenis
in 19803 and series of landmark papers later by Paulsen.6–10
The earlier papers predominantly focused on the transmission
electron microscopic features of the cilia, their distribution and
possible functions.1,3,4 Paulsen et al. proposed mechanisms of
tear outflow and acquired lacrimal obstructions, based on their
Accepted for publication March 21, 2015.
Mohammad Javed Ali receives royalties from Springer for his text book
“Principles and Practice of Lacrimal Surgery”. Other authors have no
financial or conflicts of interest to disclose.
Address correspondence and reprint requests to Mohammad Javed Ali,
F.R.C.S., Dacryology Service, L.V. Prasad Eye Institute, Hyderabad, India.
E-mail: drjaved007@gmail.com
DOI: 10.1097/IOP.0000000000000489
414 Ophthal Plast Reconstr Surg, Vol. 31, No. 5, 2015
Copyright © 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc. Unauthorized reproduction of this article is prohibited.
ultrastructural studies on functional anatomy of lacrimal sac
epithelium and cavernous body of the lacrimal sac and naso-
lacrimal duct. To the best of the authors knowledge, there is no
study that solely focussed on the SEM features of all the differ-
ent parts of a healthy lacrimal system. This study analyzed the
SEM features of the normal punctum, canaliculi, lacrimal sac,
sac–duct junction, and the nasolacrimal duct.
METHODOLOGY
Study Design and Sample Collection. Institutional review
board approval was obtained prior to the commencement of
this study. A prospective interventional study was performed
on the healthy adult lacrimal drainage systems (n = 5) obtained
from fresh exenterated specimens (3 females, 2 males, mean
age −54.4 years; range 41–63). Exenteration was performed for
malignancies unrelated to lacrimal system where preoperative
lacrimal evaluation was normal. A careful and through
dissection was carried out to isolate the entire lacrimal drainage
system from the punctum to the nasolacrimal duct. Longitudinal
and cut sections were obtained at different levels namely the
punctum, canaliculus, lacrimal sac, and nasolacrimal duct. The
ultrastructural features of the external and internal surfaces
were studied using the standard protocols of SEM.
Scanning Electron Microscopy Protocol. Scanning electron
microscopy specimens were fixed in a solution of 2.5%
glutaraldehyde in 0.1 M phosphate buffer (PBS, pH 7.2) for 24
hours at 4°C. The samples were then postfixed in 2% aqueous
osmium tetroxides for 4 hours and subjected to serial dehydration
in a series of graded alcohols. This was followed by drying
using silica vacuum desiccation for 45 minutes. The processed
samples were mounted over the stubs with double-sided carbon
conductivity tape, and a thin layer of gold coating was performed
for 3 minutes using the automated sputter coater (JEOL JFC-1600,
Tokyo, Japan). Images were taken using a JEOL-JSM 5600 SEM
(JEOL Ltd, Tokyo, Japan) at an accelerated voltage of 10 kV, and
at various magnifications between ×70 and ×10,000.
RESULTS
Punctum
Ultrastructurally, the punctum showed certain characteristic fea-
tures. End-on views showed a well-defined lumen with smooth surfaces
of punctal rim and edges (Fig. 1A). Proximal luminal surfaces were
smooth all around (Fig. 1B). Higher magnification of inner punctal sur-
faces showed a definite slightly elevated junction between the luminal sur-
faces of punctum and beginning of the vertical canaliculus (Figs. 1C–E).
Surfaces proximal to the junction were smoother while those distal to the
junction had rugae-like fine mucosal folds (Fig. 1E). Higher magnification
SEM of inner punctal surfaces showed smooth epithelium with occasional
 Ophthal Plast Reconstr Surg, Vol. 31, No. 5, 2015 Surfaces of Normal Adult Lacrimal Drainage System
FIG. 1.  Scanning electron microscopic images of the punctum.
End-on view of the punctum with central lumen and surround-
ing punctal rim (SEM ×70, A). Higher magnification clearly
showing the luminal details (SEM ×150, B). Electron micropho-
tograph showing smooth inner punctal surface with a junctional
area (J) (SEM ×350, C). Higher magnifications of another lacrimal
system showing well-defined junctional area (J). Note the PE,
smooth IP surface, and the J (SEM ×1,500, D). Still higher mag-
nification comparing the surfaces of inner punctum, junction
and the vertical canaliculus, distal to the junction (SEM ×2,500,
E). Regular surface of the canalicular epithelium with occasional
large goblet cells (SEM ×20,000, F). Ultrastructural appearance
of the punctal rims. Note the presence of throttle veins (white
arrowheads; SEM ×450, G) and a venule (black arrowhead) in a
high magnification (SEM ×5,000, H). SEM, Scanning electron
microscopy; PE, punctal edge; IP, inner punctual; J, junction.
goblet cells but no cilia (Fig. 1F). The cut surfaces of surrounding punctal
rim showed sparse and small throttle veins (Fig. 1G) with higher magnifi-
cations demonstrating venules and surrounding capillaries (Fig. 1H).
Canaliculus
The lumen was wider but variable at different portions along the
canaliculus (Fig. 2A). The external surfaces appeared rough (Fig. 2B)
with criss-cross and occasionally irregular arrangement of collagen fi-
bers (Fig. 2B, C). In comparison, the luminal or the inner surfaces were
smooth (Fig. 2D) but showed occasional valvular structures in the distal
canaliculus as large mucosal folds (Fig. 2E). The surface mucosa of the
valves was smooth, however, it demonstrated broad and blunt rugae-like
folds (Fig. 2F). Higher magnification SEM images showed a smooth epi-
thelial lining with occasional goblet cells and no cilia (Fig. 1G). Distal
segments of the canaliculus showed well-defined orbicularis muscle
© 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc. 415
Copyright © 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc. Unauthorized reproduction of this article is prohibited.
FIG. 2.  Scanning electron microscopic images of the cana-
liculus. Low magnification image showing end-on view of the
canalicular lumen (SEM ×200, A) External surface of the cana-
liculus showing rough (SEM ×200, B) surface with visible large
collagen bundles (SEM ×300, C). Smooth inner surface of the
canaliculus (SEM ×1,000, D). Lumen of the canaliculus (L) show-
ing smooth surface (S) on 1 end and valvular elevations (V) on
the other (SEM ×400, E). High magnification of the valvular area
showing the broad rugae-like mucosal folds on the surface (SEM
×1,500, F). Canalicular epithelial areas showing smooth epithe-
lium with occasional goblet cells (SEM ×25,000, G). Electron
microphotograph showing a well-defined arrangement of orbi-
cularis muscle with collagen fibres in the vicinity (SEM ×1,000,
H). SEM, Scanning electron microscopy.
fibres with numerous collagen fibres between the muscle fibres and cana-
licular surface, probably helping in attachment (Fig. 1H).
Lacrimal Sac
The external surfaces of lacrimal sac were giving a rough ap-
pearance because of collagen fibres arrangements (Fig. 3A, B). The
wall of the lacrimal sac showed numerous large capacitance vessels
and smaller arteries embedded in the collagen matrix (Fig. 3C). Very
high magnification of the cut section of the wall shows numerous
muscle fibres admixed with collagen bundles (Fig. 3D). The lumen was
wide with smooth surfaces but more prominent rugae-like structures
(Fig. 3E) and occasional large villus-like structures (Fig. 3F). Certain
areas of mucosal and submucosal elevations were well defined with
numerous scattered lymphocytes on the surface and surrounding vicin-
ity, probably representing lymphoid follicles (Fig. 3G). Very high mag-
nification showed the epithelium to be regular with numerous goblet
cells (Fig. 3H). In addition, numerous scattered glandular structures
 M. J. Ali et al. Ophthal Plast Reconstr Surg, Vol. 31, No. 5, 2015
FIG. 3.  Scanning electron microscopic images of the lacrimal
sac. Low and high magnification images showing the rough
external surfaces of the lacrimal sac (SEM ×70, A; SEM ×250, B).
Electron microphotograph of the lacrimal sac wall showing the
dense vascular plexus (SEM ×700, C). High magnification of the
sac wall showing relation of muscle bundles and collagen fibres
(SEM ×10,000, D). End-on view of the wide lacrimal sac lumen
(SEM ×300, E). Luminal surface of the lacrimal sac showing
numerous rugae-like irregular projections and occasional large
villus-like structures (SEM ×1,500, F). Electron microphotograph
showing the mucosal and submucosal well-defined elevations
possibly lymphoid follicles (SEM ×1,600, G). Glandular structure
(G) with surrounding epithelium and opening of ducts (black
arrowhead) on the epithelial surface (SEM ×25,000, H). SEM,
Scanning electron microscopy.
were noted within the epithelial surface with openings of glands in the
­vicinity, more so in the proximal areas of lacrimal sac (Fig. 3H).
Nasolacrimal Duct
The junction between lacrimal sac and nasolacrimal duct
(NLD) could reasonably be made out ultrastructurally on an external
surface examination, although not very clearly discernable (Fig. 4A).
Such a junction could also be made out on a longitudinal cut section
of the NLD (Fig. 4B) and end-on luminal views (Fig. 4C). The exter-
nal surface of the NLD showed dense collagen network with numer-
ous vascular plexus in its wall, which included wide luminal arteries
and large capacitance vessels (Fig. 4D). The lumen of the NLD was
narrower with smooth edges (Fig. 4E). The cut section of the NLD
wall showed a high density of blood vessels as compared with lac-
rimal sac wall and included arterioles, throttle veins, and large ca-
pacitance vessels (Fig. 4F). Higher magnifications show the intricate
416 © 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc.
Copyright © 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc. Unauthorized reproduction of this article is prohibited.
FIG. 4.  Scanning electron microscopic images of the nasolac-
rimal duct. Low magnification external surface image showing
the junction (J) between the lacrimal sac and NLD (SEM ×150,
A). Longitudinal cut section of the distal portion of the sac and
NLD, showing the lumen (L) of the NLD and the sac–duct junc-
tion (J) (SEM ×110, B). End-on view of the lumen (L) at the sac
duct junction (SEM ×100, C). Low magnification photograph of
the external surface of the nasolacrimal duct showing openings
of the large vessels (white arrowhead; SEM ×300, D). End-on
view of the lumen of nasolacrimal duct (SEM ×170, E). High
magnification showing vascular plexus openings embedded
in the dense matrix of large collagen bundles (SEM ×500, F).
Rugae-like internal surface of the NLD (SEM ×500, G). Electron
microphotograph showing large goblet cells with villus-like
structures on the internal surface of NLD (SEM ×3,500, H). NLD,
nasolacrimal duct; SEM, Scanning electron microscopy.
relations of the vascular plexus with the arrangement of collagen fi-
bres (Fig. 4F). The inner surfaces of NLD are smooth but irregularly
elevated like in the lacrimal sac (Fig. 4G); however, the inner sur-
face shows numerous villus-like structures, uniform epithelium with
abundance of goblet cells (Fig. 4H).
DISCUSSION
This study examined the SEM features of healthy lacrimal
drainage pathways. Distinct features of canalicular valves,
orbicularis muscles arrangement in the canalicular wall, vari-
able external and internal surface features of lacrimal sac and
nasolacrimal ducts have been elucidated uniformly in this study.
Reasonably, well-discernable junctional areas between inner
punctal surface-vertical canaliculus and lacrimal sac–NLD
points to their possible functional roles.
 Ophthal Plast Reconstr Surg, Vol. 31, No. 5, 2015 Surfaces of Normal Adult Lacrimal Drainage System
Radnot1 described the ultrastructural features of lacrimal
sac in 14 cases of dacryocystitis. Goblet cells were universally
found in all the samples, however individual or grouped cilia
were seen only in 4 specimens. Large intercellular lacunae with
disintegration of desmosomal apparatus, plasma cells and mast
cells in the interstitium and mitochondrial inclusion bodies were
noted. However, these findings were possibly secondary to the
chronic dacryocysititis and cannot be attributed to an normal
lacrimal sac.
Adenis et al.3 studied the SEM and transmission elec-
tron microscopic features of entire lacrimal systems (n = 2)
and parts of lacrimal system (n = 6) that were obtained from
cadavers. They in addition studied 2 cases of lacrimal sac and
nasal mucosa obtained from cases of dacryocystitis during a
dacryocystorhinostomy. The predominant focus of this arti-
cle was on microvilli and possible mechanism and role of its
vibratory action. The surfaces of the cells of punctum were flat
with strong intercellular junctions and sparse microvilli and
cilia. The external part of the canaliculus showed numerous
microvilli and absent cilia, whereas the internal part showed
the absence of microvilli. The lacrimal sac demonstrated mal-
phigian epithelium with scattered cilia arranged in tufts. The
cilia were found to be numerous in the nasolacrimal duct. The
current study however analyzed only healthy systems obtained
from live humans undergoing an orbital exenteration and
attempted to study all the basic ultrastructural features from
the punctum to the nasolacrimal duct.
Thale et al.6 demonstrated that the wall of the lacrimal sac
has a helical arrangement of reticular and elastic fibres of the
collagen bundles and embedded within this network of fibers is
the vascular plexus. They proposed that this arrangement allows
“wrung out” effect of the lacrimal sac during blinking, which
facilitates the tear outflow. The vascular plexus may play a cru-
cial role in drainage and absorption of the tear fluid.6,10 Paulsen
et al.8 studied the transmission electron microscopic and SEM
features of the lacrimal sac epithelium and found that the epithe-
lial cell borders were clearly distinguishable with tight junctions
and surface microvilli. In their apical portions, epithelial cells
contained large lipid droplets and secretory vacuoles. Goblet
cells were numerous but with sparse microvilli. Paulsen et al.9,10
studied the ultrastructural features of the cavernous body of the
lacrimal sac and nasolacrimal ducts and found that more than
two-third of the bony canal from the orbit to the NLD opening
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Copyright © 2015 The American Society of Ophthalmic Plastic and Reconstructive Surgery, Inc. Unauthorized reproduction of this article is prohibited.
at inferior meatus was filled by a system of wide lumened veins
and arteries. They identified 3 types of vessels in the cavernous
body; large convoluted capacitance vessels, veins with central
narrowing called as throttle veins and barrier arteries. The pro-
posed that bulging and subsiding of the cavernous body sec-
ondary to blood flow influences the opening and closing of the
lacrimal system lumens and hence regulate tear outflow. They
also proposed that possibly the mucosa in swollen state could
have been mistaken for the valves. Although such a possibility
exists, the current study has shown that at least as far as the
canaliculi are concerned, there was presence of demonstrable
valves arising from 1 wall of the canaliculus with the other walls
appearing smooth and flat.
In conclusion, ultrastructural features of external and
internal surfaces of lacrimal drainage system help in better
understanding of the tissues and their pathologies. The junctional
area between the punctum-vertical canaliculus and lacrimal sac–
NLD needs further exploration to understand their roles.
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