EP0267026A2
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EP
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Apparatus and method for cryopreparing biological tissue for ultrastructural analysis
Assignee: The University of Texas System
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Filing: 1987-11-04
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Granted: —
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Expires: 2007-11-04
Abstract
Abstract This invention relates to apparatus for the cryopreparation of biological tissue samples for ultrastructural analysis. The use of the apparatus comprises vitrifying a biological tissue sample under cryogenic temperature conditions and ultra low vacuum conditions. The depressurized, vitrified tissue sample is brought to equilibrium in a sample holder at a temperature of less than -140°C. The tissue sample is then dehydrated while maintained in a state of thermal equilibrium. After reaching equilibrium the tissue sample is optionally infiltrated with a degassed resin followed by a polymerization of the resin to form an embedded tissue sample.
Inventors
- 👤 John G. Linner
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-23 14:51
CPC Classifications
A; A01; A01N; A01N1/00; A01N1/10; A01N1/16; A01N1/168; A01N1/165; F; F26; F26B; F26B5/00; F26B5/04; F26B5/06; G; G01; G01N; G01N1/00; G01N1/28; G01N1/42
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
58 words
A method for cryopreparing a biological tissue sample for ultrastructural analysis characterized by: (a) vitrifying a biological tissue sample; (b) depressurizing the atmosphere surrounding said sample; (c) bringing said vitrified tissue sample to equilibrium at a temperature of less than -140°C; and (d) dehydrating said vitrified tissue sample while said sample is maintained in a state of equilibrium.
Claim 2
Independent Claim
58 words
A method for cryopreparing a biological tissue sample for ultrastructural analysis characterized by: (a) vitrifying a biological tissue sample; (b) depressurizing the atmosphere surrounding said sample; (c) bringing said vitrified tissue sample to equilibrium at a temperature of less than -140°C; and (d) dehydrating said vitrified tissue sample while said sample is maintained in a state of equilibrium.
Claim 3
Dependent Claim
18 words
The method of claim 1 characterized in that said vitrifying is to a temperature of -140°C or below.
Claim 4
Dependent Claim
16 words
The method of claim 2 characterized in that said vitrifying is by means of liquid nitrogen.
Claim 5
Dependent Claim
17 words
The method of claim 3 characterized in that said vitrifying is completed in less than 1 second.
Claim 6
Dependent Claim
17 words
The method of claim 1 characterized in that said vacuum is at least 3 x 10⁻⁹ mbar.
Claim 7
Dependent Claim
17 words
The method of claim 5 characterized in that said vacuum is created in less than 300 minutes.
Claim 8
Dependent Claim
21 words
The method of claim 1 characterized in that the equilibrium of said vitrified tissue sample is indicated by a constant temperature.
Claim 9
Dependent Claim
16 words
The method of claim 7 characterized in that said constant temperature is between -140°C to -196°C.
Claim 10
Dependent Claim
15 words
The method of claim 1 characterized in that said dehydration is by means of sublimation.
Claim 11
Dependent Claim
21 words
The method of claim 9 characterized in that said dehydration is enhanced by the addition of energy from a secondary source.
Claim 12
Dependent Claim
16 words
The method of claim 10 characterized in that said secondary source of energy is heat energy.
Claim 13
Dependent Claim
16 words
The method of claim 10 characterized in that said secondary source of energy is radiant energy.
Claim 14
Dependent Claim
18 words
The method of claim 12 characterized in that said radiant energy is by means of nuclear magnetic resonance.
Claim 15
Dependent Claim
22 words
The method of claim 10 characterized in that said radiant energy is infrared produced by a laser and transmitted by fiber optics.
Claim 16
Independent Claim
45 words
A method of cryopreparing biological tissue which is characterized by: (a) rapidly cooling said tissue at a rate and to a temperature sufficient to vitrify the water in said tissue; and (b) removing said water from said tissue by vaporization directly from its vitrified state.
Claim 17
Independent Claim
45 words
A method of cryopreparing biological tissue which is characterized by: (a) rapidly cooling said tissue at a rate and to a temperature sufficient to vitrify the water in said tissue; and (b) removing said water from said tissue by vaporization directly from its vitrified state.
Claim 18
Independent Claim
89 words
A method of cryopreparing biological tissue which is characterized by: (a) rapidly cooling said tissue at a rate and to a temperature sufficient to vitrify the water in said tissue; (b) subjecting said tissue containing said vitrified water to a combination of vacuum and temperature conditions at which said water is capable of vaporizing directly from its vitrified state; and (c) slowly adding energy to said tissue at a rate and in a quantity sufficient to dehydrate said tissue by vaporizing said vitrified water directly from said vitrified state.
Claim 19
Independent Claim
89 words
A method of cryopreparing biological tissue which is characterized by: (a) rapidly cooling said tissue at a rate and to a temperature sufficient to vitrify the water in said tissue; (b) subjecting said tissue containing said vitrified water to a combination of vacuum and temperature conditions at which said water is capable of vaporizing directly from its vitrified state; and (c) slowly adding energy to said tissue at a rate and in a quantity sufficient to dehydrate said tissue by vaporizing said vitrified water directly from said vitrified state.
References Cited (Backward Citations)
No citation records indexed.
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