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TOTALTEMP CRYO
MICROWAVE SAMPLE PROCESSOR
Microwave-Assisted
Freeze Substitution of Biological and Biomedical Samples
Cryo
Microwave Sample Processor with the TotalTemp Thermal Platform System specially
engineered to operate inside of the Ted Pella BioWave Microwave Oven
Description of Process:
Freeze substitution fixation (FS) of hydrated samples frozen in vitreous ice provides exceptional
preservation of structure for light and electron microscopy, and enables
immunological detection of thermo-labile antigens that otherwise are
damaged/destroyed by processing at ambient or elevated temperatures. Its use as a research tool or in clinical
pathology has, however, been limited by the relatively lengthy periods required
for passive diffusion of fixatives and organic solvents into the frozen
hydrated material.
The invention utilizes
controlled microwave (MW)
irradiation to accelerate the FS process; and comprises systems, devices and
methods for microwave-assisted processing of samples under cryo-conditions,
typically -80°C. The entire MWFS
procedure is now accomplished in less than 4 hours as compared to the
approximately 4-5 days previously required for FS process.
Description of Product:
The Cryo Microwave Sample Processor System combines two
specific technologies, controlled microwave irradiation and temperature
cycling. The controlled temperature
cycling device, a Thermal Platform (a.k.a. Hot/Cold Plate) was specially
engineered by TotalTemp using some alternative nonmetallic materials to allow
it to operate inside of a microwave environment. The Thermal Platform is cooled with Liquid Nitrogen
(L-N2) giving it a
precisely controlled temperature range of -100°C to +200°C.
The microwave oven was provided to TotalTemp by the
customer to be modified for the installation of a bulkhead interface plate to
allow the input of AC power, L-N2 and a nitrogen gas purge to prevent
excessive condensation inside the oven while running the platform at cryo
temperatures. The nitrogen gas purge is
directed inside of a clear polycarbonate cover that rests over the platform
surface. TotalTemp machined a custom
designed adapter plate with recessed pockets to support vials and petri dishes
for thermal conduction during the FS process.
The Microwave Oven is a Ted Pella Biowave Model 3451 which has special Magnetron
for continuous power from 80-750W, not pulsing power like a standard kitchen
oven.
Applications:
- Provides
superior preservation and rapid turnaround in research and high throughput
clinical laboratory settings
- Applicable
to a broad range of biological samples, hydrogels, and other hydrated
materials
- Processing
for light and electron microscopy
- Low-temperature
synthetic and analytical chemistry
Advantages:
- Reduces
processing periods from days to hours
- Improves
preservation, approaching native state
- Enables
uncomplicated, programmable operation
- Provides
excellent reproducibility with full logging and network printing
capability
- Barcode
scanning option available
Custom
Configured Adapter Plate
TotalTemp Thermal Platform
outside of oven showing custom Adapter Plate
designed to support specific
vials and petri dishes for tissue sample preparation
Freeze
Substitution Time and Temperature Chart
Time
Comparison Chart of sample preparation while under microwave irradiation (red line)
vs. sample preparation without microwave irradiation (purple line). Note
the
tremendous time difference on the “Minutes” scale.
Transmission Electron Microscope images of Bacillus Subtilis,
using ambient and cryo-preparative methods.
Mid-log
phase cultures were prepared by conventional chemical fixation (A), MW-assisted
chemical fixation (B), traditional FS (C), or by rapid FS without (D) or with
MW irradiation (E-F). Inserts show
detailed cell wall structure at a magnification equivalent to that in panel
D. Samples processed rapidly by FS
without MW irradiation exhibited poor preservation (D). Samples processed by MWFS exhibited
structures such as membranes, ribosomes, and cell wall material that were
obscured or poorly discernible by other techniques. Bars: 50 nm.
TotalTemp Technologies, Inc. 1022 West Morena Blvd., Ste H
888.712.2228 San
Diego, CA 92110
www.TotalTempTech.com