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Elect r on Micr oscopy Sciences
T ECH N I CAL DAT A SH EET
JB-4 Plus EMBEDDING MEDIA
EMS Cat. #14272-00
JB4 Plus Embedding Kit is a unique polymer embedding material that gives a higher level of morphological detail than
Paraffin processed tissues. A water-soluble media, JB-4 does not require dehydration to absolute alcohol except for
dense, bloody, or fatty tissue specimens. JB-4 Plus offers all the benefits of our regular JB-4 formulation with the
additional advantages of a cooler acting accelerator and the production of clear blocks. A clear polymer embedding
matrix is essential to numerous histological and histochemical procedures, most notably in ophthalmic applications.
A water-soluble media, JB-4 Plus does not require dehydration to absolute alcohol except for dense, bloody or fatty
tissue specimens. JB-4 is excellent for routine stains, special stains, and histochemical staining. Clearing agents such
as xylene and chloroform are not required. The polymerization of JB-4 Plus is exothermic, which is easily controlled
by polymerizing on ice or by using refrigeration at 4oC. All JB-4 Plus Type Embedding kits must be used under a
chemical fume hood.
Sections of JB-4 Plus embedded material can be cut at 0.5 to 3.0 microns or thicker. Microtones designed for plastic
sectioning are required as are glass, Ralph, or tungsten carbide, or for the best result diamond knives. Diatome U.S.
has good selection of Diamond Histo-Knives, which are excellent for sectioning JB-4 Plus blocks. Sections can be
stained for routine histological or histochemical procedures. Immunohistochemical procedures are not recommended
for JB-4 as the glycol methacrylate cannot be removed from the section and may block antigen sites for most antibody
reactions. As an alternative we recommend the LR White, Unicry or Micro-Bed (See EMS catalog for more
information)
Note: It is recommended that the Embedding Kits be used under a chemical fume hood with appropriate gloves.
Fixation: Dehydration:
Specimens can be fixed in 10% Neutral Buffered Dehydration can be completed at room temperature or
Formalin or other routine histological fixative. We 4掳C. This process can also be done with a routine tissue
suggest using HITOCHOICE鈩B庐 (EMS Cat processor, stopped at the end of the last alcohol step
#16786), a formaldehyde-free fixation. Routine and removed for infiltration. Please note that polymers
specimen sizes for soft tissue should be no more than 2 cannot be used in routine histology tissue processors, at
x 2 x 2cm with fixation at a minimum of four hours to any time. It may void the warranty and possibly begin to
overnight. Fatty or dense tissues should be fixed polymerize in the system thereby blocking the lines.
overnight. Lager bone specimens, will requires fixation Check with the manufacturer prior to attempting
overnight or longer depending on the specimen size. infiltration on any unit.
Fixation can be at room temperature or 4掳C. Cold
Infiltration:
fixation will extend the time required for the specimen
to be penetrated and fixed. Larger bone specimens will Infiltration Solution Mixing Procedures:
require longer fixation times, dependent on the size and The following amounts of material are used for one
density of the bone. Bone specimens are not decalcified 100ml batch of infiltration:
prior to fixation. Note that while JB-4 Plus can be used JB-4 Plus Solution A (Monomer) 100 ml
for bone specimens, polymerized JB-4 Plus is slightly Benzoyl Peroxide, Plasticized (Catalyst) 1.25 g
softer that the JB-4 Embedding formulation, therefore
JB-4 is recommended for most bone procedures.
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Carefully weigh 1.25 g of catalyst (benzoyl peroxide
Embedding Solution Mixing Procedure:
plasticized) and add to 100 ml Solution A while stirring
Make fresh Solution A following the directions in
on a magnetic stirrer. Mix until dissolved approximately
infiltration solution and procedures above. Do not use
10 to 20 minutes. Measurement of the catalyst is critical
old or used catalyzed Infiltration Solution for the
as it will control the rate of polymerization of the plastic
embedding solutions.
and the exothermic reaction. This infiltration solution
can be stored for up to two weeks in a dark cool area or
The following amounts of material are used for 25 ml of
in the refrigerator at 4掳C.
embedding solution:
Infiltration Procedure: Infiltration Solution 25.0 ml
Infiltration is performed at room temperature or at 4掳C. JB-4 Solution B (Accelerator) 1.0 ml (must
Do not expose the samples to heat or direct light during be an exact measurement)
EMBEDDING SOLUTION:
infiltration. The specimens should be placed in two to
three changes of infiltration solution to allow for the Embedding Solution Fresh Infiltration JB-4Plus
removal or replacement of all alcohols or tissue fluids. Volume Solution B
The amount of infiltration solution used is 50ml 50ml + 25ml
approximately 8 to 10 times that of the volume of the 25ml 25ml + 1ml
specimen. The changes of fluid should be every 10 to
90 minutes for smaller specimens. The time in each
change is dependent on the size of the specimen. When Mix 25 ml of fresh made Infiltration Solution and 1.0
infiltration is complete the tissue generally appears ml of JB-4 Plus Solution B thoroughly and begin
translucent and in most cases will sink to the bottom of embedding immediately. Drain excess Infiltration
the container. Solution off the specimen before orienting in the block
Infiltration should be done on a slow rotator, holder. The block holder is essential to exclude oxygen
hematology shaker table or inverted several times during the polymerization process. If Block holders are
during the process to allow complete saturation. not used, cover the molds with an air tight film or place
INFILTRATION SOLUTION: under vacuum at no more that 15psi, preferably in a
cold room at 4oC or a refrigerator. Beem capsules may
Infiltration Solution JB-4 Plus Benzoyl
Volume Solution A Peroxide be capped for embedding
100ml 100ml + 1.25ml
50ml 50ml + 0.625ml The blocks will be clear to very pale yellow. This color
shift is not a problem and will not effect the block
25ml 25ml + 0.3125ml
hardness. The top of the block may have a liquid film
on it and can be removed by draining or drying the
block in a desiccator for several hours to overnight.
Embedding:
The polymerization process should be under anaerobic
Deplasticizing and Staining:
conditions with the use of block holders, under light
JB-4 Plus is a glycol methacrylate based polymer and it
vacuum or in an air-tight container.
cannot be removed completely from sections, therefore
no organic solvents are required. Routine histology
Prior to mixing the Embedding Solution collect and
stains and most histochemistry can be run on the
prepare the following materials; embedding molds,
sections. High molecular weight special stains or
block holders, labels, gloves, instruments, an ice bath,
immunohistochemical reactions may not penetrate the
and the specimens. Do not pre-cool the molds as this
polymerized plastic in the sections and may result in an
may cause condensation and prevent even
incomplete reaction.
polymerization of the block face. To prevent
polymerization from occurring too fast and possible
Warning:
overheating of the tissue it is recommended that the
polymerization process for embedding be slowed by May be harmful if swallowed. Use under a fume hood
completing it in the refrigerator or in a cold room at with appropriate gloves. Components may cause
4掳C. Note that this may extend the polymerization from irritation and or allergic skin reaction. Avoid contact
several hours to overnight. with eyes, skin and clothing. Avoid inhalation of the
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vapors. Wash hands or expose areas thoroughly after
First Aid:
handling the solutions.
In case of contact with any component or mixed
Precautions: solution, immediately flush area with plenty of water
Do not heat over an open flame. Avoid electrical or for at least 15 minutes. Should either unpolymerized or
static sparks. Store un-catalyzed resin in the original polymerized material contact the eyes flush with water
containers at room temperature in a dark cool area.
Water for at least 15 minutes. Should either adjustments to the amounts recommended. The catalyst
unpolymerized or polymerized material contact the should be kept tightly sealed. The catalyst may
eyes flush with water for at least 15 minutes. If decompose with age, therefore we recommend
swallowed, drink water to excess and call a physician carefully monitoring the date received and using the
immediately. Never give anything by mouth to catalyst only with the kit it came in for best results.
someone who is unconscious.
Catalyst Disposal Procedure:
Storage: The catalyst can be destroyed by slowly adding and
mixing it in small portions of the catalyst at 4 times or
Refrigeration of the kit components is not required but
more the volume to weight of 10% sodium hydroxide
they do required storage in a cool dark place. Do not
solution in water. Do not allow material to settle in
store in the light or in a heated area as it may cause the
lumps or stand in layers and mix until dissolved
monomer to polymerize. The catalyst, plasticized
completely. Dispose of this solution, Monomer A, and
benzoyl peroxide, is organic peroxide that is shipped
the accelerator with other hazardous wastes in
dry and does not required special storage. Please note
accordance with local, state, and federal regulations.
that the catalyst is formulated to remain stable and
weigh correctly for this procedure without any
Ordering information:
Catalog # Description Size
14272-00 JB-4 Plus Embedding Kit 1 kit
14272-01 JB-4 Plus Solution A 800 ml
14272-04 JB-4 Plus Solution B 30 ml
14272-06 JB-4 Plus Catalyst 12.0 g
15740-01 Formalin Neutral Buffered 1L
64115-01 HISTOCHOICE鈩B庐 1L
70175-50 Plastic Block Holder 50/pk
70176-10 Molding Cup Tray 6 x 12 x 5mm each
70176-15 Molding Cup Tray 12 x 16 x 5mm each
70176-20 Molding Cup Tray 6 x 8 x 5 mm 3/pk
70176-25 Molding Cup Tray 2 x 15 x 5 mm 3/pk
70176-30 Molding Cup Tray 13 x 19 x 5 mm 3/pk
70180 Peel-Away Mold, Truncated Tip 8 x 8 mm 288/cs
70181 Peel-Away Mold, Truncated Tip 12 x 12 mm 288/cs
70182 Peel-Away Mold, Square Tip 22 x 22 mm 288/cs
70183 Peel-Away Mold, Rectangular Tip 22 x 30 mm 288/cs
70184 Peel-Away Mold, Rectangular Tip 22 x 40 mm 288/cs
Electron Microscopy Sciences
321 Morris Road * P.O. Box 251
Fort Washington, PA 19034
215-646-1566 * (US) 800-523-5874
Fax: 215-646-8931 * email:sgkcck@aol.com
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