Histological specimen treatment apparatus and method

Abstract

Method and apparatus for processing tissue specimens against decomposition, putrefaction and autolysis which use a simple three step procedure in a single vessel or container. First, the specimens are saturated with a solvent mixture of a ketone and a hydrocarbon, e.g., an Acetone/Hexane or an Acetone/Xylene mixture, to dissolve lipids and other cellular solutes. The container is then flooded with melted Paraffin. In a last step, the solvent mixture is vaporized and evacuated from the container, allowing the melted Paraffin to replace the vaporized solvent and impregnate the specimens. Raw, i.e., non-processed and non-burred specimens up to 5 mm thick can be processed in about 60 minutes. A solvent regenerator distills the evacuated solvent; and converts vent waste gases into carbon dioxide and water through a thermocatalytic oxidizer.

1 . A tissue processing method which comprises:
 providing a pressure and temperature controllable reaction container;   placing at least one fresh tissue specimen in said container;   treating said specimen in said container to remove solutes and replace said solutes with Paraffin, said step of treating consisting essentially of:
 introducing a temperature and pressure conditioned dissolving compound into said container; 
 allowing time for said compound to penetrate said specimen and to dilute solutes therein; 
 flooding said container with liquid Paraffin; 
 vaporizing said compound and allowing a period for said Paraffin to impregnate said specimens; 
 evacuating said compound and diluted solutes from said container; and 
 reducing the temperature in said container below the melting point of Paraffin. 
   
   
   
       2 . The method of  claim 1 , wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures. 
   
   
       3 . The method of  claim 1 , wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons. 
   
   
       4 . The method of  claim 1 , wherein said dissolving compound comprises a mixture taken from a group consisting essentially of:
 a mixture of about 60% per weight Acetone and about 40% per weight of at least one Hexane;   a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;   a mixture of about 62% per weight Acetone, about 16% per weight of at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;   a mixture of about 17% per weight Acetone, about 39% per weight of at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;   a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;   a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and,   a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.   
   
   
       5 . The method of  claim 1 , wherein said dissolving compound consists of a mixture taken from a group consisting essentially of:
 a mixture of about 60% per weight Acetone and about 40% per weight of at least one Hexane;   a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;   a mixture of about 62% per weight Acetone, about 16% per weight of at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;   a mixture of about 17% per weight Acetone, about 39% per weight of at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;   a mixture of about 46% per weight Acetone, about 25% per weight of at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;   a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and,   a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.   
   
   
       6 . The method of  claim 1 , which further comprises;
 regenerating an amount of said dissolving compound evacuated from said container in a regeneration canister.   
   
   
       7 . The method of  claim 6 , wherein said regenerating comprises:
 distilling said evacuated compound; and   converting vent waste gases into carbon dioxide and water through a thermocatalytic oxidizer.   
   
   
       8 . The method of  claim 6 , which further comprises conditioning regenerated dissolving compound for introduction into said reaction container. 
   
   
       9 . The method of  claim 8 , wherein said conditioning comprises:
 heating said dissolving compound in a solvent vessel to a temperature of approximately 60° C.; and   establishing a pressure of approximately 0.8 bars (12 psig) in said vessel.   
   
   
       10 . The method of  claim 1 , wherein said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 30 minutes. 
   
   
       11 . The method of  claim 10 , wherein said specimens are taken from a group consisting essentially of non-processed and non-burred tissue sections up to 5 mm thick. 
   
   
       12 . The method of  claim 1 , wherein:
 said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and   said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.   
   
   
       13 . The method of  claim 1 , which further comprises admitting a limited amount of Paraffin into said container before introducing said dissolving compound. 
   
   
       14 . The method of  claim 1 , wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20° C. 
   
   
       15 . The method of  claim 1 , wherein said dissolving compound comprises a lubricant for process pumps and valves. 
   
   
       16 . The method of  claim 1 , wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane. 
   
   
       17 . The method of  claim 1 , wherein said dissolving compound is vaporizable at a pressure of no more than 10 torr and a temperature of no more than about 80° C. 
   
   
       18 . The method of  claim 17 , wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20° C. 
   
   
       19 . A histoprocess which comprises:
 dissolving and removing cellular solutes in a tissue specimen using a single dissolving compound; and   replacing said solutes with Paraffin;   said process being performed in a continuous sequence of steps within a single vessel.   
   
   
       20 . The histoprocess of  claim 19 , wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures. 
   
   
       21 . The histoprocess of  claim 19 , wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons. 
   
   
       22 . The histoprocess of  claim 19 , wherein said dissolving compound comprises Acetone and at least one Hexane. 
   
   
       23 . The histoprocess of  claim 19 , wherein said steps comprise:
 saturating said specimen with said dissolving compound;   flooding said vessel with liquid Paraffin; and   vaporating said dissolving compound and said solutes to allow said Paraffin to infiltrate spaces vacated by said compound and solutes.   
   
   
       24 . The histoprocess of  claim 19 , which further comprises:
 evacuating said dissolving compound and solutes to a regeneration canister;   recuperating said dissolving compound by distillation; and   disposing of vent gases by thermocatalytic oxidation.   
   
   
       25 . The histoprocess of  claim 19 , which further comprises mixing said compound in said vessel prior to said dissolving and removing. 
   
   
       26 . The histoprocess of  claim 19 , wherein said dissolving compound comprises a lubricant for process pumps and valves. 
   
   
       27 . The histoprocess of  claim 19 , wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20° C. 
   
   
       28 . The histoprocess of  claim 19 , wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane. 
   
   
       29 . The histoprocess of  claim 19 , wherein said dissolving compound is vaporizable at a pressure of no more than 10 torr and a temperature of no more than about 80° C. 
   
   
       30 . The histoprocess of  claim 19 , wherein:
 said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and   said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.   
   
   
       31 . The histoprocess of  claim 19 , which further comprises admitting a limited amount of Paraffin into said container before introducing said dissolving compound. 
   
   
       32 . An apparatus for processing tissue specimens against decomposition, putrefaction and autolysis which comprises:
 a hermetically sealable reaction container;   means for controlling the temperature and pressure inside said container;   a source of dissolving compound connectable to said container;   a source of melted Paraffin connectable to said container; and   means for evacuating gases from said container;   wherein said dissolving compound comprises a mixture of a ketone and at least one low molecular weight hydrocarbon.   
   
   
       33 . The apparatus of  claim 32 , wherein said dissolving compound comprises a mixture taken from the group consisting of azeotropic and zeotropic dehydrant mixtures. 
   
   
       34 . The apparatus of  claim 32 , wherein said dissolving compound comprises a mixture taken from a group consisting essentially of:
 a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;   a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;   a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propano;   a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and,   a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.   
   
   
       35 . The apparatus of  claim 32 , wherein said dissolving compound consists of a mixture taken from a group consisting essentially of:
 a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;   a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;   a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propano;   a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and,   a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.   
   
   
       36 . The apparatus of  claim 32 , wherein Paraffin present in the solvent provides lubricating properties for process pumps and valves. 
   
   
       37 . The apparatus of  claim 32 , wherein said dissolving compound comprises a solvent taken from the group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons. 
   
   
       38 . The apparatus of  claim 32 , which further comprises means for regenerating vaporized dissolving compound evacuated from said container. 
   
   
       39 . The apparatus of  claim 38 , wherein said means for regenerating comprise:
 a still for recuperating said evacuated dissolving compound; and   a thermocatalytic oxidizer for converting vent waste gases into carbon dioxide and water.   
   
   
       40 . The apparatus of  claim 32 , wherein said dissolving compound has a refractive index between about 1.350 and 1.499 at 20° C. 
   
   
       41 . The apparatus of  claim 32 , wherein said dissolving compound comprises a mixture of approximately 17% to 65% per weight Acetone and 16% to 40% per weight at least one Hexane. 
   
   
       42 . The apparatus of  claim 32 , wherein:
 said time is limited to no more than approximately 30 minutes, and said period is limited to no more than approximately 50 minutes; and   said specimens comprise non-processed and non-burred tissue sections up to 10 mm thick.   
   
   
       43 . The combination of a tissue specimen with a preparation for dissolving lipids and other cellular solutes in said specimen, said preparation being taken from a group consisting essentially of:
 a mixture of about 60% per weight Acetone and about 40% per weight at least one Hexane;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO, about 20% per weight 2-Propanol and about 2% per weight Paraffin;   a mixture of about 62% per weight Acetone, about 16% per weight at least one Hexane, about 4% per weight DMSO, about 12% per weight 2-Propanol and about 6% per weight Paraffin;   a mixture of about 17% per weight Acetone, about 39% per weight at least one Hexane, about 5% per weight DMSO, about 12% per weight 2-Propanol and about 30% per weight Paraffin;   a mixture of about 46% per weight Acetone, about 25% per weight at least one Hexane, about 7% per weight DMSO and about 20% per weight 2-Propanol;   a mixture of about 53% per weight Acetone, about 26% per weight at least one Xylene, about 20% per weight Ethanol, and about 1% per weight DMSO; and,   a mixture of about 62% per weight Acetone, about 19% per weight at least one Xylene, and about 19% per weight Ethanol.   
   
   
       44 . A process for removing cellular solutes from a tissue specimen which comprises treating said specimen with a dissolving compound having a viscosity of no more than about 1.30 centipoises at 60° C. and a boiling point of more than about 80° C. at a pressure of no more than 10 torr. 
   
   
       45 . The process of  claim 44 , wherein said compound is zeotropic. 
   
   
       46 . The process of  claim 44 , wherein said dissolving compound has a viscosity of no more than about 0.75 centipoises at 60° C. and a boiling point of less than about 80° C. at a pressure of no more than 10 torr. 
   
   
       47 . The process of  claim 44 , wherein said compound is regenerated after combination with said solutes. 
   
   
       48 . The process of  claim 44 , wherein said dissolving compound is azeotropic. 
   
   
       49 . The process of  claim 44 , wherein said dissolving compound comprises a solvent taken from a group consisting essentially of ketones, esters, alcohols, aldehydes, ethers, aromatics, low molecular weight hydrocarbons, and a mixture of low molecular weight hydrocarbons. 
   
   
       50 . The process of  claim 44 , wherein said dissolving compound comprises a lubricant for process pumps and valves.