1/21/97
Two questions:
1) We would like to contact Beverly Giammara for some help and
information regarding her publications on flat molds for embedding tissue
using the microwave. If any one has any information on how to contact
her we would greatly appreciate it.
2) To any one else doing microwave embedding
We are a lab that deals with muscle and nerve samples. We would like to
use our new microwave to embed the tissue in resin (Spurr's or LR White).
We tried Beem capsules but were unable to orient the tissue properly for
sectioning with our MT2-B. I have seen reference made to the use of
flats molds in the microwave and wonder if any one has any experience or
suggestions they could share with us.
Thanks in advance
Christine Brantner and Susan Danielson
Muscle/Nerve Lab
Froedtert Hospital
Milwaukee WI
of resins: 1) Giammara's approach for flat embedding molds and 2) Giberson
and Demaree's approach for Beam Capsules.
My suggestion for flat embedding molds is to start with 50% power for 15
minutes (100% will definately give you disappointing results). Also when
using flat embedding molds, allow your blocks to cool for 15 minutes before
removing them from their molds, and vent your microwave oven during curing.
I highly recommend using an appropriately sized water load for your oven
during microwave curing in flat embedding molds (otherwise there is simply
too much energy in a microwave oven and specimen damage from over heating
will result). In addition, microwave curing in an uncalibrated microwave
oven is very tricky and usually results in disappointing results (e.g.,
incomplete curing of blocks). Simple tools that you can make and a
detailed description of how to use them to CALIBRATE YOUR MICROWAVE OVEN
are published in the The Microwave Tool Book.
When curing resins in BEEM capsules, they are placed IN a water bath-
temperature never exceeds 100 °C and curing times are ~ 75 minutes. See
the reference by Giberson RT, Demaree RS, Jr: Microwave fixation:
understanding the variables to achieve rapid reproducible results. Microsc
Res Tech 32:246, 1995
Detailed information regarding curing times of various resins in a
microwave device can be found in:
1. Giammara B. Microwave embedment for light and electron microscopy using
Epoxy resins, LR White, and other polymers. Scanning 1993;15:82-87.
2. Login GR, Dvorak AM. The Microwave Toolbook. A Practical Guide for
Microscopists (Beth Israel Hospital, Boston, 1994). The book contains a
table of curing times for resins tested.
Please contact me if you have additional questions and I will respond
directly to you.
Dr. Gary R. Login
Dept. Pathology
Beth Israel Deaconess Medical Center
330 Brookline Avenue
Boston, MA 02215
phone: 617-667-2034
fax: 617-667-8676
e-mail: glogin@bidmc.harvard.edu
A good introduction to Beverly Giammara's work with microwave embedding can
be found in Chapter 23 of The Microwave Cookbook for Microscopists, by Kok
and Boon, entitled "Microwave Exposure and Epoxy-resin Embedding for EM."
This book is available through Energy Beam Sciences.
We also have a bibliography of papers relating to this subject at our World
Wide Web site (http://www.ebsciences.com).
Using the method developed by Giammara, Spurr or LR White resin
polymerization can be done in about 30 minutes in a laboratory microwave.
Best regards,
Steven E. Slap, Vice-President
Energy Beam Sciences, Inc.
Adding Brilliance To Your Vision
ebs@ebsciences.com
http://www.ebsciences.com/