Leica Microsystems EM TP Application Note

Leica Microsystems
Brand
Leica Microsystems
Model
EM TP
Category
Microscopes
Type
Application Note

Leica Microsystems EM TP is a fully automated tissue processor designed to streamline and enhance the embedding process for electron microscopy. It offers a range of advanced features that make it an ideal solution for researchers and laboratories looking to improve the quality and efficiency of their sample preparation.

The EM TP is capable of performing all the necessary steps for embedding biological samples in epoxy resin, including postfixation, dehydration, and infiltration. It can process up to 12 coverslips at a time, making it suitable for both small and large batches of samples.

One of the key advantages of the EM TP is its ability to perform microwave fixation. Microwave fixation is a rapid and efficient method of fixation that preserves the ultrastructure of cells and tissues. It is particularly useful for samples that are sensitive to chemical fixation or that require rapid processing.

Leica Microsystems EM TP is a fully automated tissue processor designed to streamline and enhance the embedding process for electron microscopy. It offers a range of advanced features that make it an ideal solution for researchers and laboratories looking to improve the quality and efficiency of their sample preparation.

The EM TP is capable of performing all the necessary steps for embedding biological samples in epoxy resin, including postfixation, dehydration, and infiltration. It can process up to 12 coverslips at a time, making it suitable for both small and large batches of samples.

One of the key advantages of the EM TP is its ability to perform microwave fixation. Microwave fixation is a rapid and efficient method of fixation that preserves the ultrastructure of cells and tissues. It is particularly useful for samples that are sensitive to chemical fixation or that require rapid processing.

Application Note
Embedding of murine fibroblasts grown on Aclar
plastics for TEM
Material
Research
Life Science
Research
Medical
Research
Industrial
Manufacturing
Natural
Resources
related instrument Leica EM TP
2
Embedding of murine fibroblasts grown on Aclar
plastics for TEM
TISSUE CULTURE
Epithelial cells (PtK2) or fibroblasts (3T3) were grown in DMEM at 37C and 8% CO2. They were plated either
on glass coverslips, 12 mm in diameter, or on Aclar plastics (EMS, Science Services, Munich, Germany) of the
same size. The substrata were cleaned by HCl treatment for 2 h and subsequent washing in distilled water
and 70% ethanol. Before use, they were incubated in culture medium at 37°C over night.
PREFIXATION
Prefixation was performed outside of the Leica EM TP, either for 30 min with 3% glutaraldehyde in 0.1 M
cacodylate buffer, pH 7.3 at room temperature, or by microwave fixation in 0.5% glutaraldehyde, as described
in detail elsewhere (Reipert et al., 2008).
USE OF THE LEICA EM TP FOR POSTFIXATION, DEHYDRATION AND EMBEDDING IN EPOXY RESIN
Subsequent processing steps for epoxy resin embedding were performed or in an automatic tissue processor
(LEICA Microsystems, Vienna, Austria). Four baskets (large size, 3 divisions) of the tissue processor were
arranged in pairs of two at the stem assembly to give space for the uptake of 6 coverslips in vertical position
with the cells facing towards the stem. If necessary, further 6 coverslips might be added by placing them with
their back sides opposite to the coverslips already loaded.
Postfixation with 0.5% OsO4, washing in cacodylate buffer, dehydration in a series of ethanol, mediation of
the resin infiltration by propylene oxide, as well as sample infiltration in mixtures of propylene oxide and Agar
100 (Agar Scientific, Cambridge, UK) were performed according to the protocol on the right side (page 3).
Subsequently, the coverslips were taken out of the baskets for flat embedding in pure resin Agar 100 (Agar
Scientific, Cambridge, UK). Cells were infiltrated in a drop of resin for 2 h or over night. After infiltration, an
Eppendorf tube with the bottom and lid cut off was placed above the sample. After an initial heat polymeriza-
tion at 60°C for 3 h the Eppendorf tube was filled up with resin and polymerization was continued over night.
Reipert S, Kotisch H, Wysoudil B, Wiche G. (2008) Rapid Microwave Fixation of Cell Monolayers Preserves
Microtubule-associated Cell Structures. J Histochem Cytochem.
Apr 14. [Epub ahead of print]
3
LNT Application Note - EMBEDDING OF MURINE FIBROBLASTS
Fig. 1: MW-fixed and TP-embedded 3T3 fibroblasts (overview)
Bar, 50 µm
Fig. 2: MW-fixed and TP-embedded 3T3 fibroblasts (detail)
Bar, 2 µm
4
www.leica-microsystems.com
RELATED PRODUCTS
Leica EM TP
The statement by Ernst Leitz in 1907, “With the User, For the User,”
describes the fruitful collaboration with end users and driving force of
innovation at Leica Microsystems. We have developed five brand val-
ues to live up to this tradition: Pioneering, High-end Quality, Team Spir-
it, Dedication to Science, and Continuous Improvement. For us, living
up to these values means: Living up to Life.
LIFE SCIENCE DIVISION - NANO TECHNOLOGY LNT
The Leica Microsystems Nano Technology Division’s focus is to provide
the most compehensive product portfolio for the preparation of biolog-
ical, medical and industrial samples for investigation in the Electron
and Light Microscope. Excellent Sample Preparation is a prerequisite for
perfect microscopy. Your image starts here!
Leica Microsystems – an international company with a strong network
of worldwide customer services:
Active worldwide Tel. Fax
Australia ∙ North Ryde +61 2 8870 3500 2 9878 1055
Austria ∙ Vienna +43 1 486 80 50 0 1 486 80 50 30
Belgium ∙ Diegem +32 2 790 98 50 2 790 98 68
Canada ∙ Concord/Ontario +1 800 248 0123 847 405 0164
Denmark ∙ Ballerup +45 4454 0101 4454 0111
France ∙ Nanterre Cedex +33 811 000 664 1 56 05 23 23
Germany ∙ Wetzlar +49 64 41 29 40 00 64 41 29 41 55
Italy ∙ Milan +39 02 574 861 02 574 03392
Japan ∙ Tokyo +81 3 5421 2800 3 5421 2896
Korea ∙ Seoul +82 2 514 65 43 2 514 65 48
Netherlands ∙ Rijswijk +31 70 4132 100 70 4132 109
People’s Rep. of China ∙ Hong Kong +852 2564 6699 2564 4163
∙ Shanghai +86 21 6387 6606 21 6387 6698
Portugal ∙ Lisbon +351 21 388 9112 21 385 4668
Singapore +65 6550 5999 6773 0628
Spain ∙ Barcelona +34 93 494 95 30 93 494 95 32
Sweden ∙ Kista +46 8 625 45 45 8 625 45 10
Switzerland ∙ Heerbrugg +41 71 726 34 34 71 726 34 44
United Kingdom ∙ Milton Keynes +44 800 298 2344 1908 246312
USA ∙ Buffalo Grove/lllinois +1 800 248 0123 847 405 0164
Leica EM TP Application Note Embedding of murine Fibroblasts 10/2014 ∙ Copyright © by
Leica Mikrosysteme GmbH, Vienna, Austria, 2014. Subject to modifications. LEICA and the
Leica Logo are registered trademarks of Leica Microsystems IR GmbH.
  • Page 1 1
  • Page 2 2
  • Page 3 3
  • Page 4 4
  • Page 5 5

Leica Microsystems EM TP Application Note

Leica Microsystems
Brand
Leica Microsystems
Model
EM TP
Category
Microscopes
Type
Application Note

Leica Microsystems EM TP is a fully automated tissue processor designed to streamline and enhance the embedding process for electron microscopy. It offers a range of advanced features that make it an ideal solution for researchers and laboratories looking to improve the quality and efficiency of their sample preparation.

The EM TP is capable of performing all the necessary steps for embedding biological samples in epoxy resin, including postfixation, dehydration, and infiltration. It can process up to 12 coverslips at a time, making it suitable for both small and large batches of samples.

One of the key advantages of the EM TP is its ability to perform microwave fixation. Microwave fixation is a rapid and efficient method of fixation that preserves the ultrastructure of cells and tissues. It is particularly useful for samples that are sensitive to chemical fixation or that require rapid processing.

Download PDF
report

Ask a question and I''ll find the answer in the document

Finding information in a document is now easier with AI

Related papers

Other documents