visualization of microcirculation for surgery
techniques

PRODUCT DATA SHEET
Vascupaint™ Silicone Rubber Injection
Compounds

Vascupaint Kit Components•

Description
Vascupaint™ lead-free silicone injection compounds fill
and opacify the microvasculature structures for 3D ex vivo
imaging using micro-CT, dissection or brightfieldmicroscopes, or SWIR fluorescence imaging.
The injection compounds are available in two different
colors including yellow (SKU# MDL-121) for micro-CT
imaging and green (SKU# MDL-122) for NIR or SWIR
fluorescence imaging as well as clear (SKU# MDL-123) foryour custom preparations. Other colors are available uponrequest.
Vascupaint™ is a less viscous alternative to conventionallyused lead-based injection compounds making it easier toperfuse non-surviving organisms as well as human organs
for ex vivo 3D vascular cast analysis.

Features & Benefits
••

•

•

X-ray attenuation of bismuth nanoparticles inVascupaint yellow product allows for highresolution micro-CT imaging
The sub 750 nm particle size of the bismuth
particles in yellow product allows for adequate
filling of capillaries and microvasculature
Low viscosity of mixture allows for perfusion into
non-surviving organism at physiological pressuresand complete filling of microvasculature
The catalyst is a defined media that uses watermolecules to drive cross-linking of monomer. Thebottle of catalyst does not require shaking prior touse and there is no settling of cross-linker andcatalyst and the catalyst bottle does not require
shaking prior to use.

Storage
All components of this kit should be stored at room
temperature and are stable for one year from date of
manufacture

Product Safety and HandlingThis product is for R&D use only, not for drug, household,or other uses. Please review the material safety datasheet(MSDS) for proper safety and handling procedures.

Preparing Stock Solutions of Silicone andDiluent Mixtures
It is possible to prepare stock solution of the silicone anddiluent compound mixture based on the ratios used inyour experiments, for example the ratios mentioned inpoint 1 of the protocol example below. For example, you
can add 80 ml of the silicone to 100 ml of the diluent andstore for future use.

Using Previously Prepared Stock Solutions ofSilicone and Diluent Mixtures
Some ‘soft settling’ of pigments will occur in the stocksolution. You can remove this by gently inverting thebottle several times. The pigments will remain suspended
in solutions for several hours after the gentle inversion ofthe stock solution bottle.

Protocol Example

Applications•••

1 bottle of silicone (Green, Yellow, Lime, Blue, Red
or Clear), 200 ml
1 bottle of diluent, 200 ml
1 bottle of catalyst, 20 ml

Vascular corrosion casting, imaging ofangiogenesis, quantification of vascular volumes
in ischemia and tumors, microCT analysis ofvascular morphometry, assessing vasculararchitecture of organs, teaching adjunct,

A recommended starting protocol for mouse perfusions isas follows:
1.

Gently invert and/or shake the silicone bottle andthen transfer some of it’s contents to a 50 ml

5795 Av de Gaspe, Suite #228,
Montreal, Quebec, Canada, H2S 2X3
Tel: 1-514-929-9744 Fax: 1-514-360-6014, medilumine.com

conical tube. In addition, transfer some of thecontents of the diluent bottle to a 50 ml conicaltube and place both tubes in rack holder. Gentlyinvert the conical tube with silicone a few times
and then mix all kit components in a conical tube
with the following ratios:-antegrade direction for the hindlimbs or kidneys
and in retrograde direction for for brain/eye.
Two ligations can be used for in situ fixations.
(2) cut the needle tip to make it blunt before
insertion into the heart Insert the syringe needle
into heart through the left ventricle and all theway up to aorta. Cut open the right atrium toallow blood and contrast agent to flow out ofcirculation. It is also possible to cut the edges ofthe liver instead of the atrium to ensure that theoutflow of the contrast occurs less rapidly. Some
users have found that clamping away the nonneeded branches (e.g. abdominal aorta below
renal arteries in case of kidney perfusion ormesenteric vessels for perfusion of the hindlimbs or the left ventricle for the brain increases
the quality of the perfusion

4 ml silicone
5 ml diluent
0.45 ml catalyst (5% Catalyst to mixture)

Other ratios have been used depending on theperfusion procedure (retrograde or anterograde),the organ of analysis and desired stiffness ofsilicone in the vascular cast.
Transfer the mix to a petri dish for aspiration into
a 10 ml syringe with a blunt 20 or 27-gauge
needle. Make sure there are no air bubbles whenaspirating the mix.
2.

To anesthetize mouse, you can inject 100 mg/kg
ketamine with 10 mg/kg xylazine diluted in saline
via IP route for anesthesia..Make sure it iscompletely unresponsive. Then place mouse on
surgical station (tray, cork board and blue
absorbent pad), tape the mouse on the pad toprevent it from moving during injection and spray
the body of mouse with ethanol.

See example video of this procedure. Ref.
https://youtu.be/Icm1OG-vUl4
7.

If IP injection of heparin was not performed instep 2, Perfuse with heparinized PBS (5ml) first,
and then switch to a syringe, which has been filledwith formalin (5 ml) for fixation. Following fixation,
switch to the syringe with the vascupaint mixture.
Before injecting Vascupaint, put a hemostat to clipon the heart to hold the needle in place and add
crazy glue to make sure the clip is held in place
and that a back-pressure is sensed whenintroducing the mixture into the organism. By
doing so, the needle will be fixed in the aorta andleakage of Vascupaint mixture out of the heart will
be minimized, if not completely avoided. Aperfusion pump can be used to administer
Vascupaint mixture slowly to avoid any leakages
or disruptions of the hemostat clamp.

Alternatively, inject the heparin (app. 0.05 ml) IP afew minutes before or simultaneously with theanaesthetic. It is best to use more anaesthetic
than less so as to prevent coagulation. Once
heparinized, the mouse can wait for someminutes (you do not have the stress to do it asquick as possible).
3.

Cut abdominal cavity open; avoid cutting ordisturbing any internal organs.

8.

Note: avoid creating air bubbles at the tip of theneedle while switching syringes.

4.

For brain imaging procedure, gently separate theliver from the diaphragm.

9.

5.

Cut open the diaphragm to gain access to thechest cavity and unrestricted access to thedescending aorta.

Leave the mice in fridge overnight. For brain
perfusion procedures, leave the mice upside
down at 4⁰C, overnight.

6.

Two proposed methods of cannulation :(1) Cannulate the major supplying artery (in caseof mouse – it is usually aorta). Cannulate in

10. The following day, the Vascupaint inside themouse’s body should be hardened and organs
can be removed carefully. For brain procedures,the head and exposing the skull. The organ canbe further processed using standard tissue
digestion techniques for analysis with dissecting

5795 Av de Gaspe, Suite #228,
Montreal, Quebec, Canada, H2S 2X3
Tel: 1-514-929-9744 Fax: 1-514-360-6014, medilumine.com

or brightfield microscopy (*this application may
require increased level of catalyst to ensure
proper hardening of the Vascupaint). The intact
organ is ready for analysis with microCT imaging
for the Yellow version of Vascupaint and surface
vessels can be assessed with visual inspection or
microscopy to determine success of perfusion.

5795 Av de Gaspe, Suite #228,
Montreal, Quebec, Canada, H2S 2X3
Tel: 1-514-929-9744 Fax: 1-514-360-6014, medilumine.com