Home » Abstracts » Human Health Proteomics » Impact of Histological Staining of FFPE Tissues on Proteomic Analysis by Mass Spectrometry: A Comparative Study

Impact of Histological Staining of FFPE Tissues on Proteomic Analysis by Mass Spectrometry: A Comparative Study

Human Health Proteomics

Abstract #46 Posted on26 February 202512 May 2025 12 May 2025 13h28

Abstract

Proteomic analysis by mass spectrometry (MS) applied to Formalin-Fixed Paraffin-Embedded (FFPE) tissue requires precise selection of regions of interest to yield relevant data. Laser capture microdissection (LCM) is an effective approach to isolate specific cells and structures from tissue sections. However, sample preparation steps prior to LCM, specifically staining techniques, can markedly affect the quality and the quantity of proteins detected. In this study, we evaluated and compared the compatibility of different commonly used staining methods (Hematoxylin, DAB immunohistochemistry) and immuno-fluorescence methods (DAPI) with proteomic analysis by MS.

Mouse spleen sections 8µm thickness were used. Tissue surfaces of 5mm² were microdissected with the Arcturus XT. After deparaffinization and decrosslinking steps, we performed S-Trap digestion followed by timsTOF HT DIApasef MS analysis. Identification and quantification of proteins using DIANN software and comparative statistical analysis with unstained controls will determine whether the stains interact with proteins and distort the accuracy of proteomic analysis. We investigated the influence of stains on protein integrity, extraction efficiency and MS detection sensitivity after LCR. The results obtained will facilitate the selection of the most appropriate technique, ensuring an optimal balance between precise cell selection and proteomic analysis, tailored to the specific requirements of projects.

Abstract N°

Review committee

Sélectionnez le(s) utilisateur(s) chargé(s) de la revue de cet abstract.

Communication type

Speaker abstract
Oral presentation
Flash communication
Poster

Communication status

Accepted
Rejected

Final abstract N°

Note globale

Abstract content *

Max 2000 characters. Line breaks are supported.

Mouse spleen sections 8µm thickness were used. Tissue surfaces of 5mm² were microdissected with the Arcturus XT. After deparaffinization and decrosslinking steps, we performed S-Trap digestion followed by timsTOF HT DIApasef MS analysis. Identification and quantification of proteins using DIANN software and comparative statistical analysis with unstained controls will determine whether the stains interact with proteins and distort the accuracy of proteomic analysis. We investigated the influence of stains on protein integrity, extraction efficiency and MS detection sensitivity after LCR. The results obtained will facilitate the selection of the most appropriate technique, ensuring an optimal balance between precise cell selection and proteomic analysis, tailored to the specific requirements of projects.

Author(s) informations *

1	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
2	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
3	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
4	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
5	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
6	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
7	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
8	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *
1	First name * Last name * email * Presenting author * At least and only one presenting author must be selected per abstract Presenting author Do you want this email to appear in the book of abstracts? If you refuse, only the administrators and the review commitee will have access to this email as part of the review process. YesNo Laboratory * Address * ZIP code * City * Country *

Add another author

Are you an ECR? *

<7 years after last degree

Are you a Junior PI? *

If you have started your own group less than 3 years ago and are not chosen by the scientific committee for a talk, you will have the opportunity to give a keynote during an ECR parallel session

Are you an invited speaker? *

Consent *

By submitting this form I agree
- that the administrators use my personal information for the abstract revision process.
- that my name, affiliation and abstract content may be publicly displayed.

Captcha