SPI-CHEM™ SPI-PON™ 812 KIT
Use Instructions
SPI-CHEM™ / SPI-PON™ 812 KIT INSTRUCTIONS FOR USE
CAUTION:
Thoroughly read all instructions and warnings on container labels prior to use of this kit.
SPI-PON 812 resin can be used in various embedding applications in both biological and material sciences.
A cured resin block of required hardness, to match the embedded material, can be made by varying the
proportions of DDSA (softener) and NMA (hardener). It is well known that hard specimens in a soft resin
block will tear away from the block, whereas soft specimen materials will section unevenly if embedded in
a hard resin formulation. In addition to the wide range of hardness that is attainable, other advantages include
improved penetration (in biological tissue), contrast, and ease of obtaining stable thin sections.
Formulations
One or more of the following formulations can be used for most applications:
|
RESIN COMPONENT | TYPICAL MEDIUM
HARDNESS FORMULATION | TYPICAL HARD
FORMULATION |
|
SPI-PON 812 | 16.2 ml | 9.8 ml |
|
DDSA | 10.0 ml | 3.3 ml |
|
NMA | 8.9 ml | 6.9 ml |
|
DMP-30 | 0.5 ml to 0.7 ml | 0.3 ml to 0.4 ml |
Likewise, the following combined SPI-PON-Araldite formulation of Mollenhauer provides hard blocks,
good contrast and ease of sectioning.
|
RESIN COMPONENT | MOLLENHAUER'S SPI-PON-ARALDITE FORMULATION |
|
SPI-PON 812 | 10.0 ml |
|
ARALDITE 6005 | 10.0 ml |
|
DDSA | 24.0 ml |
|
DMP-30 | 0.7 ml to 0.9 ml |
1DMP-30 (accelerator) is added to either of the above formulations, with thorough mixing,
just prior to use. Additions of DMP-30 are at a 1.5 to 2.0% level (by vol.). Increasing the amount
of DMP-30 will increase "the cure rate.
MIXING OF RESIN COMPONENTS
The preparation and mixing of the resin components must be thorough and should always be performed in
a fume hood and/or a covered vessel and for a period of at least 5-10 minutes. After the DMP-30 has been
accurately measured and completely mixed into the selected resin, the complete formulation can be used
to impregnate and/or embed the sample material.
Avoid contact of the mixture with moisture. The mixture, without DMP-30 is stable for 6 months at 4°C, or
for several days at room temperature. When refrigerated, the mixture, in its closed container must be
allowed to equilibrate to room temperature before use, to prevent condensation of moisture.
SAMPLE PREPARATION
Sample for resin infiltration and embedding should be dry. Biological samples, fixed and dehydrated through
absolute alcohol are further dehydrated in two 15 minute changes of propylene oxide. Infiltration is accomplished
using the complete formulation and propylene oxide (1:1), while gently agitating for at least 1 hour. Infiltration
of the specimen is continued in at least one change of the complete formulation for 1 hour.
Final embedding of the sample is accomplished by placing the specimen in aliquot of the complete formulation,
located in appropriate flat or capsular embedding molds.
Hardening or polymerization of the above resins can be accomplished following curing in an oven at 60°C
for 24-48 hours, after which time the blocks are allowed to cool to room temperature for subsequent ultramicrotomy.
REFERENCES
1. Luft, J.H.J. Biolphs. Biochem, Cytol. 9, 409 (1961)
2. Mollenhauer, H.H., Stain Technology 39 11 (1964)
DISCLAIMER
The information given in this bulletin is to the best of our knowledge accurate, but no warranty is expressed
or implied. It is the user's responsibility to determine the suitability for his own use of the products described
herein; and since conditions of use are beyond our control, we disclaim all liability with respect to the use of
any material supplied by us. Nothing contained herein shall be construed as a recommendation to use any product
or to practice any process in violation of any law or any government regulation.
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Friday May 16, 2008
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