EP0220269A1 - Method of impregnating a tissue specimen with paraffin - Google Patents
Method of impregnating a tissue specimen with paraffinInfo
- Publication number
- EP0220269A1 EP0220269A1 EP19860902936 EP86902936A EP0220269A1 EP 0220269 A1 EP0220269 A1 EP 0220269A1 EP 19860902936 EP19860902936 EP 19860902936 EP 86902936 A EP86902936 A EP 86902936A EP 0220269 A1 EP0220269 A1 EP 0220269A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- bath
- pressure
- paraffin
- tissue specimen
- tissue sample
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 239000012188 paraffin wax Substances 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 13
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229940057995 liquid paraffin Drugs 0.000 claims abstract description 5
- 239000012024 dehydrating agents Substances 0.000 abstract description 10
- 239000000470 constituent Substances 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 30
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 230000001744 histochemical effect Effects 0.000 description 2
- 238000005470 impregnation Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/305—Fixative compositions
Definitions
- the invention refers to a method of impregnating a tissue specimen with paraffin.
- tissue specimens are trea with a dehydrating agent, alcohol and acetone, respective before the tissue specimens are immersed in a bath of liquid paraffin.
- the pressure of the bath is lowered so muc that the respective dehydrating agent is vaporized and thereby evaporates from the tissue specimens as well-as from the paraffin bath, the pressure of the bath then being increased again to atmospheric pressure.
- An object of the invention is to provide a method of impregnation that eliminates the disadvantages of the methods known hitherto.
- Another object of the invention is to provide a method of impregnation in which the treatment with a dehydrating agent can be eliminated.
- the tissue specimen is immersed, either directly after an optional treatment of the tissue specimen with a fixing agent or after the tissue specimen optionally has been treated with a fixing agent and/or a dehydrating agent, in a bath of liquid paraffin which is kept below a predetermined temperature at which thermolabile component and properties of the tissue specimen undergo changes, and that thereafter, the pressure of the bath is lowered from atmospheric pressure to a predetermined pressure, at which water and/or, when appropriate, the dehydrating agent vaporizes. Then the pressure of the bath is increased again to atmospheric pressure.
- a tissue specimen When carrying out a histochemical diagnosis, normally a tissue specimen is first treated with a fixing agent, usually formalin, and thereafter with a dehydrating agent, e.g. methanol, ethanol or acetone, before the tissue specimen is immersed in a bath of liquid paraffin. In this bath the tissue specimen is impregnated with paraffin. The tissue specimen impregnated in this way with paraffin is then embedded in a paraffin block which can be cut by means of a microtome knife to obtain thin cuts (1-5 urn thic which can thereafter be examined in a microscope after the paraffin has been removed and after dyeing.
- a fixing agent usually formalin
- a dehydrating agent e.g. methanol, ethanol or acetone
- the tissue specimen is directly immersed into the paraffin bath.
- a paraffin having a melting point of about 37-44 C has been chosen and the bath is kept at a temperature of about 47 C at the most, i.e. below the temperature at which it has been observed that thermolabile components and properties of the tissue specimen begin to change. It should be pointed out that this temperature can vary somewhat depending on which type of tissue specimen one wants to examine.
- the pressure of the paraffin bath is thereafter lowered from atmospheric pressure to at least about 76 mm Hg, i.e. the pressure at which water boils at a temperature of about 46 C.
- the pressure of the bath is increased again to atomospheric pressure.
- the lowering of the pressure as well as the pressure increase must not take place too abrupt.
- the tissue material and cells present therein can burst through sudden pressure changes during the pressure lowering phase and can be compressed during the pressure increasing phase.
- the tissue specimen can also be treated with a dehydrating agent, for example methanol, ethanol or acetone, before the sample is immersed into the paraffin bath which also in this case is kept below the temperature at which thermolabile components and properties of the tissue specimen are changed.
- a dehydrating agent for example methanol, ethanol or acetone
- the pressure of the paraffin bath has to be lowered to at least about 510 mm Hg, i.e» the pressure at which acetone vaporizes at a temperature of about 45 C.
- the acetone will vaporize and evaporate from the tissue specimen as well as from the paraffin bath.
- the pressure of the bath must also in this case be lowered to at least 76 mm Hg.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE8501959 | 1985-04-23 | ||
| SE8501959A SE8501959L (sv) | 1985-04-23 | 1985-04-23 | Framstellning av ett vevnadspreparat |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP0220269A1 true EP0220269A1 (en) | 1987-05-06 |
Family
ID=20359937
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP19860902936 Withdrawn EP0220269A1 (en) | 1985-04-23 | 1986-04-22 | Method of impregnating a tissue specimen with paraffin |
Country Status (3)
| Country | Link |
|---|---|
| EP (1) | EP0220269A1 (sv) |
| SE (1) | SE8501959L (sv) |
| WO (1) | WO1986006479A1 (sv) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5089288A (en) * | 1989-06-24 | 1992-02-18 | Berger Hermann J | Method for impregnating tissue samples in paraffin |
| DE69600484T2 (de) * | 1996-08-02 | 1999-04-01 | Milestone Srl | Verfahren zur Bearbeitung von organischen Proben |
| DK1005633T3 (da) * | 1997-08-20 | 2008-12-15 | Univ Miami | Vævsfikserings-dehydratiserings-fedtfjernelses-imprægneringsmetode med höj kvalitet og kontinuerligt gennemlöb |
| US6793890B2 (en) | 1997-08-20 | 2004-09-21 | The University Of Miami | Rapid tissue processor |
| ATE551606T1 (de) * | 2001-10-01 | 2012-04-15 | Leica Biosystems Melbourne Pty | Gewebeprozessor zur behandlung histologischer gewebepräparate |
| US8554372B2 (en) | 2003-09-29 | 2013-10-08 | Leica Biosystems Melbourne Pty Ltd | System and method for histological tissue specimen processing |
| US20090298172A1 (en) | 2008-05-28 | 2009-12-03 | Steven Paul Wheeler | Histological specimen treatment apparatus and method |
| US10365189B2 (en) | 2015-05-07 | 2019-07-30 | Steven Wheeler | Histological specimen treatment |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2932112A1 (de) * | 1979-08-08 | 1981-02-26 | Carlos Prof Dr Thomas | Verfahren und vorrichtung zur einbettung von entwaesserten gewebeproben in paraffin |
-
1985
- 1985-04-23 SE SE8501959A patent/SE8501959L/sv not_active Application Discontinuation
-
1986
- 1986-04-22 EP EP19860902936 patent/EP0220269A1/en not_active Withdrawn
- 1986-04-22 WO PCT/SE1986/000185 patent/WO1986006479A1/en unknown
Non-Patent Citations (1)
| Title |
|---|
| See references of WO8606479A1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| SE8501959L (sv) | 1986-10-24 |
| WO1986006479A1 (en) | 1986-11-06 |
| SE8501959D0 (sv) | 1985-04-23 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
| AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AT BE CH DE FR GB IT LI LU NL SE |
|
| STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
| 18D | Application deemed to be withdrawn |
Effective date: 19870125 |