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ES2540120B2 - Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions - Google Patents
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ES2540120B2 - Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions - Google Patents

Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions Download PDF

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Publication number
ES2540120B2
ES2540120B2 ES201300524A ES201300524A ES2540120B2 ES 2540120 B2 ES2540120 B2 ES 2540120B2 ES 201300524 A ES201300524 A ES 201300524A ES 201300524 A ES201300524 A ES 201300524A ES 2540120 B2 ES2540120 B2 ES 2540120B2
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ES
Spain
Prior art keywords
location
procedure
molecular markers
microscopic dimensions
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.)
Active
Application number
ES201300524A
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Spanish (es)
Other versions
ES2540120A1 (en
Inventor
Rosa Ana PICAZO GONZÁLEZ
María De Los Ángeles SÁNCHEZ PÉREZ
Belén SÁNCHEZ MALDONADO
María Teresa ENCINAS CEREZO
Juan Antonio GILABERT SANTOS
José María ROS RODRÍGUEZ
María Pilar GARCÍA PALENCIA
Carmen Belén MARTÍNEZ-MADRID
Pedro ARANDA ESPINOSA
Paola Stefanía TINETTI PINTO
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universidad Complutense de Madrid
Original Assignee
Universidad Complutense de Madrid
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Universidad Complutense de Madrid filed Critical Universidad Complutense de Madrid
Priority to ES201300524A priority Critical patent/ES2540120B2/en
Priority to PCT/ES2014/000089 priority patent/WO2014191590A1/en
Publication of ES2540120A1 publication Critical patent/ES2540120A1/en
Application granted granted Critical
Publication of ES2540120B2 publication Critical patent/ES2540120B2/en
Active legal-status Critical Current
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Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/36Embedding or analogous mounting of samples
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31Apparatus therefor

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Optics & Photonics (AREA)
  • Biophysics (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Procedimiento de doble inclusión para el estudio histológico y/o de localización de marcadores moleculares en especímenes pluricelulares de dimensiones microscópicas.#La presente invención se refiere a un procedimiento de doble inclusión de especímenes pluricelulares de dimensiones microscópicas (70-300 {mi}m) que permite mantener la disposición espacial tridimensional de las muestras para, posteriormente, realizar estudios histológicos, inmunohistoquímicos y/o de localización de marcadores moleculares. El procedimiento incluye la preparación de la muestra mediante lavados con solución tamponada; la fijación de la muestra durante 10-15 minutos, a temperatura de 2-8° C; la deshidratación de la muestra con concentraciones crecientes de etanol durante 10-15 minutos cada una, a una temperatura de 2-8° C, la preinclusión en agar y la inclusión en parafina, prescindiendo de la centrifugación de la muestra en todas las etapas del procedimiento. Así mismo, la invención incluye un kit con los elementos necesarios para desarrollar el procedimiento descrito en la invención.Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions. # The present invention relates to a double inclusion procedure of multicellular specimens of microscopic dimensions (70-300 m) which allows the three-dimensional spatial arrangement of the samples to be maintained to subsequently perform histological, immunohistochemical and / or location studies of molecular markers. The procedure includes sample preparation by washing with buffered solution; fixation of the sample for 10-15 minutes, at a temperature of 2-8 ° C; dehydration of the sample with increasing concentrations of ethanol for 10-15 minutes each, at a temperature of 2-8 ° C, preinclusion in agar and inclusion in paraffin, regardless of centrifugation of the sample at all stages of the process. Likewise, the invention includes a kit with the necessary elements to develop the procedure described in the invention.

Description

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Claims (1)

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ES201300524A 2013-05-31 2013-05-31 Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions Active ES2540120B2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
ES201300524A ES2540120B2 (en) 2013-05-31 2013-05-31 Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions
PCT/ES2014/000089 WO2014191590A1 (en) 2013-05-31 2014-05-30 Double-inclusion method for the study of the histology and/or localisation of molecular markers in pluricellular specimens of microscopic dimensions

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
ES201300524A ES2540120B2 (en) 2013-05-31 2013-05-31 Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions

Publications (2)

Publication Number Publication Date
ES2540120A1 ES2540120A1 (en) 2015-07-08
ES2540120B2 true ES2540120B2 (en) 2015-10-27

Family

ID=51988051

Family Applications (1)

Application Number Title Priority Date Filing Date
ES201300524A Active ES2540120B2 (en) 2013-05-31 2013-05-31 Double inclusion procedure for histological study and / or location of molecular markers in multicellular specimens of microscopic dimensions

Country Status (2)

Country Link
ES (1) ES2540120B2 (en)
WO (1) WO2014191590A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111426532A (en) * 2020-04-09 2020-07-17 北京大学深圳医院 Method for making bacterial wax blocks and bacterial wax blocks as a method for positive quality control of acid-fast staining

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2458365B1 (en) * 2010-11-24 2019-04-10 Milestone S.r.l. A two-step method and system for cold formalin fixation of organic tissue samples

Also Published As

Publication number Publication date
ES2540120A1 (en) 2015-07-08
WO2014191590A1 (en) 2014-12-04

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