SE2350857A1 - A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysis - Google Patents
A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysisInfo
- Publication number
- SE2350857A1 SE2350857A1 SE2350857A SE2350857A SE2350857A1 SE 2350857 A1 SE2350857 A1 SE 2350857A1 SE 2350857 A SE2350857 A SE 2350857A SE 2350857 A SE2350857 A SE 2350857A SE 2350857 A1 SE2350857 A1 SE 2350857A1
- Authority
- SE
- Sweden
- Prior art keywords
- sample
- optionally
- treating
- concentration
- enzyme
- Prior art date
Links
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 title abstract 6
- 206010028980 Neoplasm Diseases 0.000 title abstract 3
- 238000000034 method Methods 0.000 title abstract 3
- 239000012188 paraffin wax Substances 0.000 title abstract 2
- 238000002360 preparation method Methods 0.000 title abstract 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract 3
- 102000004190 Enzymes Human genes 0.000 abstract 3
- 108090000790 Enzymes Proteins 0.000 abstract 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 abstract 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 abstract 3
- 229940088598 enzyme Drugs 0.000 abstract 3
- VHJLVAABSRFDPM-UHFFFAOYSA-N 1,4-dithiothreitol Chemical compound SCC(O)C(O)CS VHJLVAABSRFDPM-UHFFFAOYSA-N 0.000 abstract 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 abstract 2
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 abstract 2
- 102000057297 Pepsin A Human genes 0.000 abstract 2
- 108090000284 Pepsin A Proteins 0.000 abstract 2
- 229940071106 ethylenediaminetetraacetate Drugs 0.000 abstract 2
- 238000002509 fluorescent in situ hybridization Methods 0.000 abstract 2
- 238000010438 heat treatment Methods 0.000 abstract 2
- 229940111202 pepsin Drugs 0.000 abstract 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 abstract 1
- 102000035195 Peptidases Human genes 0.000 abstract 1
- 108091005804 Peptidases Proteins 0.000 abstract 1
- 239000002253 acid Substances 0.000 abstract 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6841—In situ hybridisation
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The disclosure relates to a method for the preparation of one or more formalin fixed paraffin embedded (FFPE) tumor tissue sample(s) for use in a subsequent analysis, such as in fluorescent in-situ hybridization, comprising the steps of: (a) deparaffinizing the sample; (b) heat treating the sample by adding citrate and exposing the sample to an increased temperature of about 80 °C at a duration of about 45 to 120 minutes; (c) optionally permeabilizing the sample; (d) optionally acid treating the sample; (e) enzyme treating the sample, by adding at least one proteolytic enzyme to the sample; (f) optionally formamide treating the sample; (g) optionally re-fixating the sample; (h) hybridizing the sample; wherein the method further comprises at least one additional treatment chosen from the group comprising: (A) an additional step of heat treatment performed as part of step (b), comprising adding glycerol at a concentration of about 20% to the sample and exposing the sample to an increased temperature in the interval of about 90-95 °C at a duration of about 10-30 minutes; (B) an additional step of heat treatment performed as part of step (b), comprising adding dithiotreitol (DTT) and ethylene diamine tetraacetate (EDTA) to the sample and exposing the sample to an increased temperature of about 80 °C; (C) an enzyme treatment performed as part of step (e), wherein pepsin is added at a concentration of about 0,05 mg/ml in combination with at least one additional enzyme having the ability to increase the effective concentration of pepsin; and (D) a refixation performed as part of step (g), comprising an alkaline treatment in order to stabilize the sample, followed by (i) a first fixation by adding methanol and acetic acid, and (i) a second fixation by adding formalin at a concentration of about 1%; thereby obtaining a tumor tissue sample having an essentially intact morphology prepared for subsequent FISH analysis.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE2350857A SE2350857A1 (en) | 2023-07-06 | 2023-07-06 | A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysis |
| PCT/SE2024/050651 WO2025010012A1 (en) | 2023-07-06 | 2024-06-28 | A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE2350857A SE2350857A1 (en) | 2023-07-06 | 2023-07-06 | A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SE2350857A1 true SE2350857A1 (en) | 2025-01-07 |
Family
ID=91856295
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SE2350857A SE2350857A1 (en) | 2023-07-06 | 2023-07-06 | A method for the preparation of one or more formalin fixed paraffin embedded tumor tissue samples for use in a subsequent analysis |
Country Status (2)
| Country | Link |
|---|---|
| SE (1) | SE2350857A1 (en) |
| WO (1) | WO2025010012A1 (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011104027A1 (en) * | 2010-02-26 | 2011-09-01 | Qiagen Gmbh | Process for parallel isolation and/or purification of rna and dna |
| WO2013046033A1 (en) * | 2011-09-30 | 2013-04-04 | Dako Denmark A/S | Hybridization compositions and methods using formamide |
| WO2021037869A1 (en) * | 2019-08-28 | 2021-03-04 | Ventana Medical Systems, Inc. | Assessing antigen retrieval and target retrieval progression quantitation with vibrational spectroscopy |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TW201346037A (en) | 2012-05-04 | 2013-11-16 | Chang Gung Medical Foundationlinkou Chang Gung Memorial Hospital | Specimen pretreatment method for reducing fluorescence auto-interference in fluorescence in-situ hybridization |
| WO2016190236A1 (en) * | 2015-05-28 | 2016-12-01 | コニカミノルタ株式会社 | Probe reagent for in-situ hybridization |
| CN105018598B (en) | 2015-06-05 | 2018-06-26 | 厦门艾德生物医药科技有限公司 | The FISH preprocess methods and pretreatment fluid of a kind of FFPE samples |
| CN107828862A (en) | 2017-12-15 | 2018-03-23 | 郑州大学第附属医院 | The preprocess method of lymphoma tissue specimens paraffin embedding slices fluorescence in situ hybridization detection |
| CN115261368A (en) | 2022-08-16 | 2022-11-01 | 北京思尔成生物技术有限公司 | Ready-to-use pepsin working solution and preparation method and application thereof |
-
2023
- 2023-07-06 SE SE2350857A patent/SE2350857A1/en unknown
-
2024
- 2024-06-28 WO PCT/SE2024/050651 patent/WO2025010012A1/en unknown
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011104027A1 (en) * | 2010-02-26 | 2011-09-01 | Qiagen Gmbh | Process for parallel isolation and/or purification of rna and dna |
| WO2013046033A1 (en) * | 2011-09-30 | 2013-04-04 | Dako Denmark A/S | Hybridization compositions and methods using formamide |
| WO2021037869A1 (en) * | 2019-08-28 | 2021-03-04 | Ventana Medical Systems, Inc. | Assessing antigen retrieval and target retrieval progression quantitation with vibrational spectroscopy |
Non-Patent Citations (3)
| Title |
|---|
| Abdul Rauf Shakoori, T.A. Bhat, A.A. Wani (eds.), Chromosome structure and aberrations, 2017, pp. 343-367, DOI 10.1007/978-81-322-3673-3_16 * |
| Chin S-F et al., "A simple and reliable pretreatment protocol facilitates fluorescent in situ hybridisation on tissue microarrays of paraffin wax embedded tumour samples", J Clin Pathol: Mol Pathol., 2003, Vol. 56, pp. 275-279 * |
| Ensinger C et al., "Improved technique for investigations on archival formalin-fixed, paraffin-embedded tumors by interphase in-situ hybridisation", Anticancer research, 1997, Vol. 17, pp. 4633-4638 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2025010012A1 (en) | 2025-01-09 |
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