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Technovit® 8100 Glycol Methacrylate is a HEMA-based plastic-embedding system for studies with light microscopy. It is suitable for embedding all tissues in medicine, zoology and botany. Sections of decalcified or briefly decalcified iliac crest biopsies and implanted biomaterials can be used for more than just histological staining; they can also be used for enzyme chemistry and immunohistochemistry.
Technovit® 8100 is a combination of a practically odourless plasticizer and a hydrophillic plastic. Technovit® 8100 was specifically developed for cold polymerization (+4°C).
Note: While hardening the embedding form must be hermetically sealed because the polymerization system is oxygen-sensitive.
Prepare Technovit® 8100 in accordance with the step-by-step instructions. Place the fixated and dehydrated specimens in the infiltration solution. A low temperature and agitation of the specimens is beneficial during the entire embedding process.
Prepare the polymerization mixture according to the instructions and then fill the embedding cavities. Position the infiltrated specimens therein. Hermetically cover the cavities with films. Place on a pre-cooled gel plate or thin layer of ice at 4°C to harden.
The films are removed after polymerization is complete and blocked with Histobloc® and Technovit® 3040.
It is not possible to elute the plastic before staining or reaction.
Designation | Quantity |
Technovit® 8100 | 1 x 500 ml Basic Solution 5 x 0,6g hardener 1 1 x 30g hardener 2 1 x 500 pcs PE films |
Color | Transparent |
Density = spec. weight g/cm3 (DIN 53479) |
1,08 |
Refractive index Monomer Polymer |
1,4485 1,4990 |
Storage temperature | max. 25°C |
Shelf life | 2 years |
The following instructions for fixation and dehydration are not necessarily required. Technovit® can also be infiltrated and polymerized after other pre-treatment.
Airtight glass or PE disposable containers (approx. 20 ml) must be used for the entire process!
Tip: The specimens must be constantly agitated during fixation, dehydration and infiltration!
In order to achieve optimal immunohistochemical results, it is recommended to work at 4°C throughout the entire embedding process and to aim for short fixation times. Fix the smallest possible pieces of tissue (1mm thickness) in 2% paraformaldehyde in phosphate buffer pH 7.4 at 4°C for 3-4 hours. Subsequently, retreat for 12 hours (overnight) in phosphate buffer pH 7.4 with an additional 6.8% sucrose at 4°C.
Dehydrate the tissue in cold acetone 100% for at least one hour at 4°C. Change as often as possible in the first minutes until the acetone remains clear.
Making the infiltration solution: | |
Technovit® 8100 Basic Solution | 100 ml |
+ Technovit® 8100 hardener 1 | 1 bag, 0,6 g |
Dissolve in a clean, detergent-free PE or glass container and then place at 4°C. When sealed, the infiltration solution is stable for a maximum of four weeks at 4°C.
Transfer the specimen directly from the acetone to the pre-cooled infiltration solution. The specimens remain therein for 6-10 hours at 4°C.
Making the Polymerization solution: | |
Infiltration solution, 4°C | 15 ml |
+ Technovit® 8100 hardener 2 cooled | 0,5 ml |
Measure with standard pipetting aids and mix well in a PE or glass container. Then, carefully mix the infiltrated specimen in a sealed container for approx. five minutes.
The color of the polymerization solution changes first to yellow-green, but after hardening it becomes colorless.
Completely fill the Histoform cavities with a disposable pipette, position the tissue therein and immediately cover with transparent PE film. Multiple films can be used for a cavity in order to hermetically seal the cavity. Do not press out bubbles; rather, apply more polymerization solution and add new film. During polymerization (at least 3 hours) the embedding form must be placed on a cooling plate or thin layer of ice at 4°C. Do not let the form or specimens come into contact with moisture.
Histoform Q | ||||||
Material | Room temp. approx. +20°C |
Refrigerator +4°C | Refrigerator on ice 0°C | |||
Technovit® 8100 30:1 |
-- | 69 | 48 | |||
Technovit® 8100 35:1 |
-- | 52 | 42 | |||
Technovit® 8100 40:1 |
-- | 50 | 41 |
Histoform S | ||||||
Material | Room temp. approx. +20°C |
Refrigerator +4°C | Refrigerator on ice 0°C | |||
Technovit® 8100 30:1 |
69 | 21 | 12 | |||
Technovit® 8100 35:1 |
67 | 19 | 11 | |||
Technovit® 8100 40:1 |
65 | -- | -- |
Remove the film at room temperature with tweezers once hardening is complete.
The specimens are blocked with Histobloc® and Technovit® 3040 so that they can be removed from the PTFE mold. Store blocks that are not needed immediately (for immunohistochemistry) at a cool temperature in plastic bags or similar.
One obtains the best cutting results with a rotation microtome, with the Technovit® Histoblade in combination with the Heraeus knife holder or a hard metal knife (glass diamond knife). Tightly clamp the blocks in the totem cam system on the microtome. Dryly remove the sections with forceps and place in a bath (Aqua dest.). Place directly on a coated object holder and let dry for 2 hours or more at 37°C. Dry sections that are not needed immediately (for immunohistochemistry) at room temperature and store for a maximum of five days at 4°C.
PolymerizationOverview of how to make the solution | ||||||
Solution | Ethanol | Basic solution Technovit® 8100 |
Hardener 1 Technovit® 8100 |
Infiltration Solution | Hardener 2 Technovit® 8100 |
Application-temp |
Infiltration | 100 ml | 0,6 (1 bag) | 4°C |
For example, submerse the object holder in a solution of 0.5% alcian blue (8GXL Sigma) at 65°C for 15 minutes or liquidly coat the object holder with 0.1% poly-L-lysine (Sigma). All standard coated object holders may be used.
The sections must dry for at least two hours at 37°C. Place the non-deplasticized sections directly in the stain solution or start with enzymatic pre-treatment.
Determination of immuno factors is possible with AP, PAAP, APAAP, ABC, avidin-biotin, streptavidin and immunofluorescence methods.
The use of wetting agent, e. g. Tween, in the rinsing buffer is discouraged. The peroxidase should be dissolved in buffer.
With the constantly changing range of new products for histochemistry and immunohistochemistry, is is always advisable to follow the respective manufacturer instructions.
Heraeus Kulzer, 2014