Journal of Anatomy

J. Anat. (2016) 229, pp582--590 doi: 10.1111/joa.12504

A comparative study of vascular injection fluids in

fresh-frozen and embalmed human cadaver forearms
D. E. Doomernik,1 R. R. Kruse,2 M. M. P. J. Reijnen,3 T. L. Kozicz1 and J. G. M. Kooloos1
1
Department of Anatomy, Radboud University Medical Center, Nijmegen, the Netherlands
2
Department of Surgery, Isala Clinics, Zwolle, the Netherlands
3
Department of Surgery, Rijnstate Hospital, Arnhem, the Netherlands

Abstract
Over the years, various vascular injection products have been developed to facilitate anatomical dissections.
This study aimed to compare the most commonly used vascular injection products in fresh-frozen and formalin-
embalmed cadaver specimens. An overview of the properties, advantages and limitations of each substance was
given, and a comparison of vascular infusion procedures in both preservation methods was made. A literature
search was performed in order to identify the most commonly used vascular injection products. Acrylic paint,
latex, gelatin, silicone, Araldite F and Batson’s No. 17 were selected for the study. One fresh-frozen and one
embalmed cadaver forearm were infused with each injection product according to a uniform protocol. The
curing time, skin- and subcutaneous tissue penetration, degree of filling of the arterial tree, extravasations,
consistency of the injected vessels during dissection, and the costs of each injection fluid were noted. There
was a large variation between the injection fluids in processing- and curing time, colour intensity, flexibility,
fragility, elasticity, strength, toxicity and costs. All fluids were suitable for infusion. The penetration of injection
fluid into the skin and subcutaneous tissue was significantly better in fresh-frozen specimens (P = 0.002 and P =
0.009, respectively), with significantly smaller branches casted (P = 0.004). Vascular infusion of fresh-frozen
cadaver specimens results in a significantly better filled coloured arterial tree, enabling more detail to be
achieved and smaller branches casted. The biomechanical properties of fresh-frozen soft tissues are less affected
compared with formalin fixation. All the injection fluids studied are suitable for vascular infusion, but their
different properties ensure that certain products and procedures are more suitable for specific study purposes.
Key words: acrylic resin; blood vessels; cadaver; epoxy resin; gelatine; latex; silicone; vascular anatomy; vascular
infusion; vascular injection fluids.
developing structures dates back to the 15th century, when
Introduction
Leonardo daVinci (1452–1519) made wax casts and glass
Anatomical dissections have been extensively used for many models of the chambers of the heart to study the blood cir-
years and have made a major contribution to the knowl- culation (Hodde & Nowell, 1980). Jan Swammerdam (1637–
edge of vascular anatomy. The first study in which the term 1680) is usually regarded as the inventor, but he replicated
‘arteries’ was used to describe pulsating blood vessels dates daVinci’s method by injecting melted wax, mixed with fats
back to 340 BC (Herophilos). In 1628, William Harvey (1578– and stained with cinnabar, into arteries and veins to study
1657) introduced the idea of a blood circulation, which was capillaries of the lung (Whitten, 1928; Hodde & Nowell,
confirmed by Marcello Malpihgi (1628–1694) and Antonie 1980; Paweletz, 2001).
van Leeuwenhoek (1632–1723) with the discovery of capil- Over the past decades, many different vascular injection
laries in 1661 and 1668, respectively (Paweletz, 2001). products have been utilised to facilitate dissection of blood
The use of injection products to study the vascular pat- vessels. Jean Riolan (1580–1657) introduced coloured dyes
tern of normal and pathological organs, tissues and to demonstrate the branched pattern of the vascular system
(Paweletz, 2001). In addition, Voigt (1809–1890) used gela-
tin as an injection medium and Grossner (1901) added
Correspondence
Indian ink as a pigment to enhance the contrast of gelatin
D. E. Doomernik, Department of Anatomy, Radboud University
Medical Center, Internal postal code 109, Postbox 9101, 6500 HB (Hodde & Nowell, 1980). With the introduction of
Nijmegen, the Netherlands. T: +31 (24) 3613341; E: Denise. roentgenography (1895), contrast media, such as chalk,
Doomernik@radboudumc.nl mercury, barium sulphate and lead oxide, were added to a
Accepted for publication 10 May 2016 suspending medium such as gelatin, oil or latex (Bergeron
Article published online 22 June 2016 et al. 2006). Other substances, such as metal, celloidin,

© 2016 Anatomical Society

 Vascular injection fluids, D. E. Doomernik et al. 583

rubber, silicone, plastoid, resins and polyurethane elastomer
(PU4ii), have been more recently developed as vascular
injection products (Whitten, 1928; Bergeron et al. 2006;
Meyer et al. 2007).
A lot of information can be found on vascular injection
products for anatomic dissections, such as Spalteholz clear-
ing and maceration or corrosion casting in embalmed,
fresh and fresh-frozen human and animal cadaver speci-
mens. However, there is little uniformity concerning the
use of vascular injection products and infusion techniques.
To the best of the authors’ knowledge, no studies have
been published that compare more than two different
vascular injection products using a uniform protocol. In
addition, no studies were found in which the results of
vascular injections and the preservation method were
compared.
The present study was designed as a pivotal study in
preparation of cadaver studies on the collateral circulation
of the lower limb using vascular infusion techniques. The
aim of the study was twofold. Firstly, it was wanted to
assess the differences between embalmed and fresh-frozen
cadaver specimens, in order to visualise blood vessels up to
1 mm in diameter. Second, it was aimed to find the most
suitable vascular injection product for vascular infusion and
dissection. To answer these questions, the literature was
this study could facilitate other researchers in selecting a
suitable injection fluid and preservation method for a
particular study purpose.
Materials and methods
Design
An anatomical cadaver study was performed at the Department of
Anatomy of the Radboud University Medical Center (Radboudumc),
Nijmegen, the Netherlands. The cadaver specimens were obtained
according to the Dutch Body Donation Program for Science and
Education (Wet op de lijkbezorging, 1991). Body donations of
humans aged 60 years and older with a valid handwritten testa-
ment were accepted in the study.
Cadaver specimens
Six fresh-frozen and six embalmed intact cadaver forearms with no
signs of trauma, vascular pathology or (surgical) scars were included
for analysis with an average age of 81 years (range 65–93 years).
Four embalmed and one fresh-frozen cadaver specimens were
excluded and replaced because of excessive leakage of injection
fluid, distal branching of the brachial artery (BA), or absence of an
intact palmar arch. Cadaver characteristics, such as age, sex, cause
of death and storage time were noted (Table 1).

reviewed to find the most commonly used vascular injection

products. In a uniform design, the six most commonly used
vascular injection fluids were compared in fresh-frozen and
formalin-embalmed cadaver specimens. In this study, the
properties of the casting products have been summarized,
that is the degree of filling of the arterial tree, venous
return, extravasations, colour intensity, flexibility, fragility,
elasticity, strength, advantages and limitations of the
selected vascular injection fluids were given. The results of
Embalmment
The cadaver specimens were embalmed within 72 h after death
through the right femoral artery (or vein, in any case of arterioscle-
rosis). For embalming, a mixture consisting of 25 mL phenol, 330 g
sodium chloride, 300 g chloral hydrate 98% crystallised, 1 L formalin
37%, 500 mL ethanol 100%, 300 mL pure glycerol with a specific
gravity of 1.26, warm tap-water up to a total volume of 5 L and 5 L
cold tap-water was prepared, according to local protocol. The 10 L
embalming fluid was infused in 1.5–2.0 h at a constant gradient of

Table 1 Cadaver specimen characteristics.

Number Group Side Infusion fluid Age Sex Cause of death Storage

1 Embalmed Left Acrylic paint 79 Female Unknown 7 years, 10 months

2 Embalmed Left Latex 88 Male Unknown 6 years, 10 months
3 Embalmed Left Gelatin 65 Male Cerebro-vascular accident 3 years, 6 months
4 Embalmed Right Silicone 93 Female Pneumonia 1 year, 10 months
5 Embalmed Right Araldite F 68 Female Metastatic disease of pancreas carcinoma 14 years
6 Embalmed Left Batson’s No. 17 87 Female Gasto-intestinal bleeding 1 year, 10 months
7 Fresh-frozen Right Acrylic paint 81 Male Respiratory insufficiency 6 months
8 Fresh-frozen Right Gelatin 85 Female Cardiac failure 2 years, 10 months
9 Fresh-frozen Right Latex 78 Male Metastatic disease of pulmonary cancer 10 months
10 Fresh-frozen Right Silicone 78 Female Euthanasia 2 years, 2 months
11 Fresh-frozen Left Araldite F 91 Male Cardiac failure 6 months
12 Fresh-frozen Left Batson’s No. 17 78 Male Alzheimer’s disease 1 year, 1 month
13 Embalmed Right – 79 Female Unknown 7 years, 10 months
14 Embalmed Right – 86 Female Unknown 6 years, 10 months
15 Embalmed Right – ? Unknown Unknown > 15 years
16 Embalmed Right – 88 Male Exhaustion 2 years, 6 months
17 Fresh-frozen Right – 79 Male Cerebro-vascular accident 2 years, 9 months

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584 Vascular injection fluids, D. E. Doomernik et al.

2.0 m water column (19.6 kPa) and stored at room temperature for Table 2 Composition of the selected vascular injection products.
24 h. After 24 h, 210 mL embalming fluid was injected intramuscu-
larly in both upper- and lower extremities. The embalmed speci- Composition
mens were stored for at least 6 months in a 5% formalin solution at Category Selected item of injection fluid Amount
room temperature. Before use, the 5% formalin was rinsed out
with cold tap-water and the cadaver specimens were stored in a Dye Acrylic paint1 Acrylic paint 100 mL
2.5–3% formalin solution at room temperature. Water 100 mL
Gelatin Gelatin powder2 Gelatin powder 10 g (10%)
Freeze Water 100 mL
The fresh-frozen cadaver specimens were frozen within 72 h after Acrylic paint1 5 mL
death at 40 °C for at least 1 day and stored at 20 °C. Before use, Latex Liquid latex3 Latex 100 mL
the cadaver specimens were thawed at room temperature for at Water 7.5 mL
least 24 h. Pigment4 7.5 g
Silicone Silicone5 Polymer PR-10 95 mL
Cross-linker CR-20 5 mL
Selection of the injection fluids Catalyst CT-32 3 mL
Pigment4 1g
A literature search was performed to identify the most commonly Epoxy Araldite6 Araldite F 20 mL
used vascular injection fluids. Reports on the application of vascular resin Dilutioner DY 026 SP 60 mL
injection fluids in human cadaver or animal studies, published Hardener HY 2967 45 mL
between 1 January 1939 and 1 November 2013, in the PubMed, Pigment4 1.2 g
MEDLINE, EMBASE and Cochrane databases were selected. The fol- Acrylic Batson’s No. 177 Monomer Base 100 mL
lowing MeSH terms were used: blood vessels, injections, cadaver, resin solution A
human, corrosion casting, latex, gelatin, silicone, resin, acrylic resin, Catalyst 12 mL
epoxy resin, silicone elastomers, colouring agents, elastomers, poly- Promoter 12 drops
urethane and ink. Other non-MeSH terms, such as vascular injec- Pigment 3%
tion, vascular infusion, vascular casts, injection replica, corrosion
cast, latex, gelatin, silicone, Araldite, Batson’s, Technovit, Mercox, 1
Gallery, ADVEO Belgium NV, Deinze, Belgium.
methylmethacrylate, acrylic paint, Indian ink, dyes and PU4ii, were 2
Boom B.V., Meppel, the Netherlands.
also used. These terms were applied in various combinations in 3
Wilsor Kunstharsen, Biddinghuizen, the Netherlands.
addition to the use of the ‘related articles’ and ‘citing articles’ func- 4
VOF Verfmolen de Kat, Zaandam, the Netherlands.
tion. Articles were selected on the basis of title and abstract. Full- 5
Dow Corning Corporation, Midland, MI, USA.
text articles were studied with restriction of language of publica- 6
Huntsman Corporation, The Woodlands, TX, USA.
tion (English). Non peer-reviewed reports or articles of which no full 7
Polysciences Inc., Warrington, PA, USA.
text electronic or paper version was available in the university
library were excluded. No manual cross-referencing was performed.
excluded and replaced. The arterial system was irrigated with 500
mL hand-warm (37 °C) tap-water and 10 mL soap solution (Biotexâ;
Composition of the injection fluids Unilever, Rotterdam, the Netherlands) for removing blood clots
(Lametschwandtner et al. 1984; Sanan et al. 1999; Alvernia et al.
Six categories of injection fluids were identified for vascular infu-
2010), lowering surface tension (Van der Zwan & Hillen, 1990) and
sion, that is dye, gelatin, latex, silicone, epoxy resins and acrylic
increasing flexibility of the specimen (personal communication from
resins. From each category, one injection fluid was selected,
W.J.A. van Wolferen). The irrigation time was noted with a maxi-
depending upon the availability, the possibility of shipping the pro-
mum of 90 min and the process was repeated after 24 h (Sanan
duct to the Netherlands, the ability to order small quantities and
et al. 1999).
the price per quantity. The selected injection fluids and their com-
positions are summarised in Table 2. Irrigation was performed with an infusion system, which is
schematically represented in Fig. 1 (Whitten, 1928; Hanstede & Ger-
rits, 1982; Van der Zwan & Hillen, 1990). The pressure required for
Vascular infusion protocol infusion was obtained from a compressed air line and regulated
with a reducer (A). The pressure on the manometer of the reducer
Cannulation of the radial (RA) and ulnar arteries (UA) (A) resembled the pressure (P) in a 1000 mL glass bottle (B, off-label)
The BA was identified and dissected up to the branch of the RA and was kept at a constant gradient of 2 m water column (19.6 kPa;
and the division in the UA and interosseous artery. Each vessel was Hodde & Nowell, 1980).
isolated 1–1.5 cm from the surrounding tissue and cannulated with
curved blunt needles (Van Straten Medical B.V., Nieuwegein, the Coloured injection of the arterial system
Netherlands). The needles were inserted 2–3 cm into the vessels and
The apparatus in Fig. 1 was used for coloured injection of the arte-
secured with 2-0 silk sutures to prevent dislocation and leakage.
rial system of the cadaver forearms. A constant pressure of 2 m ‘in-
fusion fluid’ column, that is the pressure needed for the infusion
Irrigation of the arterial system fluid to reach the top of a 2 m column, was used to ensure a rela-
The patency of the palmar arch was tested manually by flushing 60 tive constant and comparable flow for each infusion fluid. The infu-
mL hand-warm (37 °C) tap-water through the UA until backflow sion was started in the UA and continued until backflow from the
from the RA occurred. In case of lack of patency the specimen was RA occurred. Following this, the cannula in the RA was closed and

© 2016 Anatomical Society

 Vascular injection fluids, D. E. Doomernik et al. 585

leakage of the infusion fluid from other vessels was prevented by
placing vessel clamps. Infusion was stopped by closing the UA when
the fluid level in the bottle (C) no longer decreased and reached a
stable state for at least 5 min. The infusion time was noted with a
maximum of 90 min. The specimens were cured at room tempera-
ture (20 °C) for at least 24 h and the curing process was evaluated
daily for up to 1 week.
For gelatin infusion, the infusion system was slightly modified.
The glass bottle (C) was placed in a warm water bath ( 50 °C), the
connecting tube between (C) and (D) was isolated with aluminium
foil, and the cadaver specimens were placed in a sink with warm
tap-water ( 50 °C). The gelatin mixture was heated above 60 °C
and continuously measured with a thermometer to prevent early
solidification of the gelatin mixture (< 50 °C).
For each infusion, 200 mL mixture was prepared and the process-
ing time was recorded. All injection products were liquids and of
low viscosity, however, no quantitative viscosity measurements
were performed. Preparation of the silicone, Araldite and Batson’s
No. 17 mixtures (Table 2) was performed in a fumigation hood,
because of the toxic vapours. All experimenters wore protective
clothes and gloves. In cases of silicone, Araldite and Batson’s No. 17
infusion, additional respiratory filters were used.
Qualitative and quantitative assessment of the arterial
system
Dissection was performed after complete curing of the infusion
fluid using a uniform protocol. The skin and subcutaneous tissue
were removed and assessed for penetration of the injection fluid
under a stereomicroscope with 40 9 magnification (Wild M650;
Leica Microsystems, Wetzlar, Germany). All coloured vessels were
counted and the diameters of the smallest vessels were measured
with a digital calliper (Mahr, 16ES, resolution 0.01 mm; Hanstede &
Gerrits, 1986). Then, each compartment of the forearm and hand
were dissected, with special attention paid to the degree of filling
of the arterial tree, venous return, extravasations and the consis-
tency of the injected vessels during dissection, that is colour inten-
sity, flexibility, fragility, elasticity, strength. The definitions of these
properties that were used are summarised in Table 3. Additional
notes about the advantages and limitations of each technique were
made.

Data analysis
Data were noted and analysed using SPSS© for Windows© version
20.0 (SPSS, Chicago, IL, USA). A Mann–Whitney U-test of signifi-
cance was used to compare the penetration of the injection fluids
and the vessels size in fresh-frozen and embalmed cadaver speci-
mens. For all tests, P < 0.05 was considered statistically significant.
The Pearson product-moment correlation coefficient (r) was used to
assess the correlation between vessel diameter measurements of
individual raters. A value of r ≥ 0.70 was considered acceptable.

Fig. 1 Infusion system for irrigation and coloured injection of the

arterial system. The pressure needed for infusion is obtained from a
compressed air line and regulated with a reducer (A). The pressure on
the manometer of the reducer (A) resembles the pressure (P; SI- unit
kPa) in a 1000 mL glass bottle (B, off-label). The glass bottle (C) con-
tains the irrigation or coloured injection fluid, and (D) resembles the
cannula in the UA.
Results
Literature search
A total of 1011 articles was found using the search strategy
outlined, 738 articles were available in full text format and
divided into six categories of injection products for vascular

Table 3 Definitions.

Item Range Definition

Colour intensity Bright (++) to dull ( ) A substance is bright when it is clearly visible in the large and smaller vessels
Flexibility Flexible (++) to stiff ( ) A substance is flexible when it is easy to manipulate and behaves
like the surrounding tissues during manipulation
Fragility Brittle (++) to solid ( ) A substance is brittle when damaged by minimal tissue manipulation
Elasticity Elastic (++) to rigid ( ) A substance is elastic when it recoils in its original shape after manipulation
Strength Solid (++) to soft ( ) A substance is solid when it is resistant to changes in volume and shape
Toxicity Toxic (++) to harmless ( ) A substance is toxic when specific precautions must be taken, such as
working in a fumigation hood, or protective cloths and respiratory
filters must be worn

This table supports the interpretation of the results presented in Table 6.

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586 Vascular injection fluids, D. E. Doomernik et al.

infusion, that is dye, gelatin, latex, silicone, epoxy resins
and acrylic resins (Table 4).
Fresh-frozen vs. embalmed specimens
Water irrigation of the embalmed group to wash out blood
clots was completed in 5–15 min. The fresh-frozen speci-
mens were irrigated in < 2 min. The infusion time of the
injection products in the embalmed specimens varied
between 20 and 90 min, compared to between 7 and 11
min in the fresh-frozen group. The penetration of the injec-
tion products into the vessels of the skin and subcutaneous
tissue was significantly better in fresh-frozen specimens (P
= 0.002 and P = 0.009, respectively; Fig. 2A). In addition, sig-
nificantly smaller coloured vessels were seen in the fresh-
frozen group compared with the embalmed group (P =
0.004; Table 5). A strong correlation of r = 0.946 was seen
between the measurements of individual raters.
enhanced injection and filling of small peripheral vessels by
up to 0.35 mm in embalmed and 0.04 mm in fresh-frozen
cadavers. Acrylic paint does not cure at room temperature,
which complicates dissection through leakage into the sur-
rounding tissues when vessel walls are damaged (Fig. 2B;
Tables 3 and 6).
Gelatin
Gelatin is a water-soluble, commonly available and cheap
product, with a temperature-dependent viscosity. Fluid
gelatin at a temperature of 60 °C enhanced complete filling
of the arterial tree up to 0.03 mm in embalmed and < 0.01
mm in fresh-frozen cadavers. The gelatin dissolved due to
formalin preservation in the embalmed cadavers and due
to water vaporisation in the fresh-frozen cadavers. This was
mainly seen in the distal vessels, where it lost its colour
intensity during dissection (Tables 3 and 6).

Evaluation of the injection products
Acrylic paint
Acrylic paint is a water-soluble, commonly available and
cheap product, with a high viscosity and high colour inten-
sity. Dilution with tap-water lowered the viscosity and
Latex
Latex is a cheap, water-soluble, flexible and elastic product.
The latex used in this study had a relatively high viscosity
and had to be diluted with tap-water before infusion. This
dilution caused longer curing times at room temperature
(Fig. 2C; Tables 3 and 6).

Silicone
The silicone used in this study is a very expensive and toxic
Table 4 Results of systematic literature search.
product, however, this does not apply to all silicone. Sili-
Total number Full text Excluded
cone resulted in a minimal colour intensity, because the pig-
Category of articles articles articles ment did not completely dissolve. However, no micro
embolisms were observed under microscopic evaluation of
Dye1 84 64 20 the skin and subcutaneous tissue, and complete filling of
Gelatin2 64 59 5 the arterial tree was observed in both the embalmed and
Latex3 219 193 26 fresh-frozen groups (Tables 3 and 6).
Silicone4 50 41 9
Epoxy resins5 16 15 1
Araldite
Acrylic resins6 564 352 212
Other7 14 14 0 Araldite is a strong, but fragile epoxy resin, because it
Total 1011 738 273 lacks elasticity. In smaller structures (< 1 mm) in particu-
lar, minimal manipulation can easily cause structures to
1
Includes that is (Indian) ink, methylene blue, acrylic paint and break down. Araldite F has a high viscosity with a ‘syr-
other non-specified dyes. upy’ texture, which makes it hard to measure and apply
2
Includes gelatin mixtures of varying concentrations (3–25%) in smaller amounts. Dilution using a dilutioner is an
with and without the addition of dyes and contrast media.
3
essential element in lowering the viscosity of the resin
Includes that is Ward’s Natural Science coloured latex, Thiel’s
to facilitate infusion. An exothermic reaction was seen
DGM 85, Type AL 1337 Dunlop, Mouldtex, Plasti Dip, neoprene
and other non-specified coloured latex. after adding the Hardener HY 2967 to the Araldite F/
4
Includes 3110 RTV and 3481 RTV silicone rubber (Dow Corning), Dilutioner DY 026 SP mixture, which resulted in melting
MV-130 Microfil and other non-specified silicone. of a plastic stirrer and a three-way valve. No macroscopic
5
Includes Araldite F, Araldite CY223, Araldite CY221 and other damage to the cadaver specimens and vessel walls was
non-specified epoxy resins. observed due to this thermal reaction (Fig. 2D; Tables 3
6
Includes Mercox, Technovit, Batson’s No. 17, MMA and other
and 6).
non-specified acrylic resins.
7
Includes PU4ii, polyurethane foam, PMC-780, polyester resin,
platogen G, vicrylic resin, tensol cement No. 70, latex resin and Batson’s No. 17
other non-specified materials. Waxes were excluded from the Batson’s No. 17 is a strong, colour-intense, but fragile
search. and relatively expensive, acrylic resin. The lack of

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 Vascular injection fluids, D. E. Doomernik et al. 587

A B

C D

Fig. 2 Coloured injection of fresh-frozen

cadaver specimens. (A) Penetration of
coloured injection fluid in the subcutaneous
tissue. (B) Extravasations of acrylic paint. (C)
Latex infusion. (D) Silicone infusion with less
colour-intense parts caused by non-dissolving
pigment. (E) Araldite infusion.

Table 5 Number and smallest diameter of the coloured vessels in skin and subcutaneous tissue.

Skin Subcutaneous tissue Smallest diameter

Embalmed (n) Fresh-frozen (n) Embalmed (n) Fresh-frozen (n) Embalmed (mm) Fresh-Frozen (mm)

Acrylic paint 80 155 0 89 0.35 0.04

Gelatin 50 346 17 266 0.16 < 0.01
Latex 1 349 19 202 0.1 0.03
Silicone 12 183 9 106 0.22 < 0.01
Araldite F 111 523 124 298 0.03 < 0.01
Batson’s No. 17 22 22 37 1319 0.08 < 0.01
Mean ( SD) 46.0  42.8 1217.2  222.3 34.3  45.6 380.0  467.5 0.16  0.13 0.018  0.014
P-value 0.002 0.009 0.004

elasticity complicates dissection of smaller vessels, because Discussion

it easily breaks. Its toxicity complicates infusion, as a
large fumigation hood or an additional room with spe-
cial ventilation and respiratory masks are required. An
exothermic reaction during polymerisation was described
by the producer, however, no thermal reaction was
observed during preparation, infusion and dissection
(Tables 3 and 6).
This is the first study in which six different vascular injection
fluids have been compared in a uniform design in both
fresh-frozen and embalmed cadaver specimens. The results
have shown that all injection fluids are suitable for vascular
infusion, and indicate that fresh-frozen cadavers provide a
significant advantage. All fluids had their own specific

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588 Vascular injection fluids, D. E. Doomernik et al.

Table 6 Summary of the properties of the different vascular injection fluids.

Processing Hardening
time (min, time (h, Filling of the Venous Colour Costs3
20 °C) 20 °C) arterial tree return Extravasations1 intensity2 Flexibility2 Fragility2 Elasticity2 Strength2 Toxicity2 (100 mL)

Acrylic Unlimited –4 Partial No Multiple ++ ++ n.a.4 € 0.14

paint
Gelatine Unlimited > <1 Complete No Single +/ + +/ + +/ € 0.30
60 °C
Latex Unlimited 48–72 Partial No Several ++ ++ ++ +/ +/ € 1.66
Silicone < 60 24–48 Complete No No +/ +/ + + + ++ € 117.09
Araldite F 45 4–6 Complete No No + ++ ++ ++ € 5.40
Batson’s 30–45 3–4 Complete No Single ++ ++ ++ ++ € 27.35
No. 17

n.a., not assessable.

1
Extravasations were only seen in embalmed specimens.
2
The definitions of these items are summarized in Table 4.
3
Price per 100 mL, excluding taxes and shipping costs.
4
Acrylic paint was the only injection fluid that did not cure.

characteristics, as was also found by Peng et al. (2009), who
compared three vascular injection products (red latex perfu-
sion, ink for diaphanisation and histology, and gelatin–lead
oxide for radiography) to reveal the blood supply of the
brachial plexus and its main branches in fresh cadavers.
They concluded that each injection method has specific
characteristics, making them more or less suitable in the
study of the extrinsic- and intrinsic blood supplies to the
nerve.
There are several possible explanations for the significant
advantage of fresh-frozen over embalmed cadaver speci-
mens for vascular infusion. It is known that formalin fixa-
tion changes the biomechanical properties of soft tissue by
cross-linking the proteoglycan monomers (denaturalisation
of proteins), which results in blood clotting and stiff-, rigid-,
brittle- and ‘difficult-to-handle’ tissues (Viidik & Lewin,
1966; Chapman et al. 1990). In the formalin-embalmed
group, rigid tissues and solid blood clots, irrigation prob-
lems (no backflow and leakage), longer infusion times, little
stained small vessels and extravasations were encountered.
These problems were probably related to both the formalin
itself and formalin fixation. The stiffness of soft tissue and
the high vascular resistance due to formalin fixation influ-
enced the distal filling of the arterial tree. Furthermore, the
formation of blood clots by formalin itself might have
caused obstruction or distal emboli, which resulted in
incomplete filling. In the fresh-frozen group, none of these
problems occurred, despite the freezing, and, thus,
explained the significantly better stained and more intri-
cately filled arterial tree. However, embalmed cadavers also
offer some benefits over fresh-frozen cadavers, such as lim-
ited exposure to pathogens, durability and long preserva-
tion time, which enable long-term dissection (Eisma et al.
2011).
In the current study, acrylic paint was the only injection
fluid that did not cure, so that the acrylic paint leaked into
the surrounding tissues in the case of damaged vessel walls
during dissection. In contrast, Maga et al. (2013) reported
no hardening or leakage problems with acrylic paint emul-
sion (Liqutiex R; Binney and Smith, Crayola LLC, Northamp-
ton County, PA, USA) in brains, stored in a 10%
formaldehyde solution for 2 weeks, followed by sectioning
in slices of 1–4 mm. In addition, Hupkens et al. (2010, 2013)
described a way to harden acrylic paint by leaving the limb
in dry air for 1 day, followed by freezing for the next 2 days
and thawing it for 1 or 2 days.
Latex was used by more than 200 authors in various con-
centrations. For example, El-Barrany et al. (1999) reached
vessel diameters of 0.6 mm in preserved cadavers with an
unspecified latex solution, and Zenn & Heitmann (2003)
reached vessel diameters of 1.1 mm in fresh cadavers with a
low viscosity mixture of undiluted 66% latex and 7%
ammonium hydroxide, this cured in 24 h at room tempera-
ture. In contrast, this study reached penetration into vessels
of < 0.1 mm in both embalmed and fresh-frozen cadavers,
using a high-viscosity 33% latex and < 1% ammonium
hydroxide mixture, diluted with water, with a polymerisa-
tion time of 48–72 h. This long polymerisation time is unfa-
vourable for fresh-frozen cadaver specimens, because of
the accelerated decay of the specimens at room tempera-
ture. Therefore, a low-viscosity latex with short polymerisa-
tion times would be favourable if the fresh cadaver
specimens are required for scientific research or educational
purposes. In addition, a limitation of the available literature
is that all studies report the use of latex as an injection fluid
to answer a specific research question on vascular anatomy,
but do not report the technical aspects and specific charac-
teristics of the latex used (Tables 3 and 6). This study is the

© 2016 Anatomical Society


first that reports the technical aspects of a specific latex,
therefore it is recommended that researchers always test a
specific latex in a pilot experiment, as there are so many
types of latex with varying properties available.
To the authors’ knowledge, the silicone PR-10 polymer
(Dow Corning, Midland, MI, USA) is used only for plastina-
tion of cadaver specimens (Baker, 1999) and not for vascular
infusion. Therefore, the authors performed a pilot study
(unpublished results) in which different concentrations of
polymer, cross-linker and catalyst were combined. The com-
bination of 95 mL Polymer PR-10, 5 mL Cross-linker CR-20, 3
mL catalyst CT-32 and 1 g was found to be the most accept-
able combination. However, the optimal mixture for vascu-
lar infusion has yet to be determined. Another widely-used
silicone for vascular infusion of cadaver specimens supplied
by Dow Corning is the RTV 3110 silicone (Sanan et al. 1999).
Unfortunately, the RTV 3110 silicone could not be shipped
to the Netherlands in small amounts, which rendered it too
expensive for inclusion in the study.
Araldite was found to penetrate into vessels < 0.01 mm.
Similar results were observed by Van der Zwan & Hillen
(1990), who used Araldite F to study cerebral vascularisation
in dogs. Approximately 90% of the arterioles with a diame-
ter of 15 lm were filled with a smallest vessel diameter of
5–10 lm. In contrast to the current results, venous return of
Araldite was observed (Van der Zwan & Hillen, 1990). Poly-
merisation is an exothermic reaction, which could theoreti-
cally cause damage to vessel walls and surrounding tissues.
However, consistent with the literature studied, no macro-
scopic soft tissue damage was observed (Hanstede & Gerrits,
1982; Van der Zwan & Hillen, 1990).
This study shows that all six vascular injection products
are suitable to facilitate dissection. However, the study is
limited by the small number of cadaver specimens in each
group, especially in the description of properties, limitations
and advantages of the different injection fluids. In addition,
whilst describing the product properties, no data were col-
lected if a product struggled to harden or cure in a moist
environment, or when the product had been in contact
with the flushing tap-water. Further sample selection bias
was found in the individual variability of the cadaver speci-
mens, who differed in age, duration of formalin fixation or
freezing, diameter of blood vessels and vascular resistance.
Despite cadaver variability, a strong correlation of r = 0.946
was observed between the measurements of individual
raters, which supports the accuracy of the measurements. In
addition, some equivalent substitutes of injection fluids not
previously described in literature were used, which makes it
difficult to compare with other studies.
This study only focused on vascular injection of the arte-
rial system of the forearm. Therefore, special considerations
should be made before using these materials in other
research projects. Further research should focus on vascular
injection of other hollow-organ systems, such as the
venous- or lymphatic systems, and on complementing the
© 2016 Anatomical Society
Vascular injection fluids, D. E. Doomernik et al. 589
overview of properties of the different infusion fluids with
factors, such as adhesion to vessel wall, deformation of the
cast, shrinkage and viscosity, which were not included in
this report.
Conclusion
In conclusion, all the injection fluids studied are suitable for
vascular infusion in both fresh-frozen and embalmed cada-
ver specimens. Vascular infusion of fresh-frozen cadaver
specimens resulted in a significantly better stained and
detailed coloured arterial tree, because the biomechanical
properties of soft tissues were less affected compared with
the formalin-fixated tissues. However, fresh-frozen cadaver
specimens decay faster, therefore, the use of embalmed
cadavers or embalming of fresh-frozen cadavers are valu-
able alternatives when long-term dissection is required.
The selection of a suitable vascular infusion fluid should
depend on the specific study purpose. Latex is preferable
when flexibility and elasticity are required for dissection.
Gelatin is an alternative, but its use is limited by the process-
ing temperature of 60 °C. Silicone offers elasticity and
strength, however, less expensive alternatives are desirable
and available. When a strong polymer is required, Araldite
or Batson’s No. 17 present the best options.
Acknowledgements
The authors would like to thank Marjan Doom and colleagues from
the Department of Morphology, Faculty of Veterinary Medicine,
Ghent University, Belgium for their helpful advice regarding the
study protocol. Furthermore, the authors would like to thank B.C.J.
van den Bosch, resident at Radboudumc Nijmegen, the Nether-
lands, for his help in collecting data.
Author contributions
Study conception and design: Doomernik, Kruse, Reijnen,
Kooloos. Acquisition of data: Doomernik and Kruse. Analy-
sis and interpretation of data: Doomernik, Kruse, Kooloos.
Drafting of the manuscript: Doomernik. Critical revision of
the manuscript: Doomernik, Kruse, Reijnen, Kozicz, Koo-
loos. Approval of the article: Doomernik, Kruse, Reijnen,
Kozicz, Kooloos.
Conflicts of interests
None.
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