Detailed Information on Publication Record
2019
Drug Penetration Analysis in 3D Cell Cultures Using Fiducial-Based Semiautomatic Coregistration of MALDI MSI and Immunofluorescence Images
MACHÁLKOVÁ, Markéta, Barbora PAVLATOVSKÁ, Jan MICHÁLEK, Adam PRUŠKA, Karel ŠTĚPKA et. al.Basic information
Original name
Drug Penetration Analysis in 3D Cell Cultures Using Fiducial-Based Semiautomatic Coregistration of MALDI MSI and Immunofluorescence Images
Authors
MACHÁLKOVÁ, Markéta (203 Czech Republic, belonging to the institution), Barbora PAVLATOVSKÁ (203 Czech Republic, belonging to the institution), Jan MICHÁLEK (203 Czech Republic, belonging to the institution), Adam PRUŠKA (203 Czech Republic, belonging to the institution), Karel ŠTĚPKA (203 Czech Republic, belonging to the institution), Tereza NEČASOVÁ (203 Czech Republic, belonging to the institution), Katarzyna Anna RADASZKIEWICZ (616 Poland, belonging to the institution), Michal KOZUBEK (203 Czech Republic, belonging to the institution), Jan ŠMARDA (203 Czech Republic, belonging to the institution), Jan PREISLER (203 Czech Republic, belonging to the institution) and Jarmila NAVRÁTILOVÁ (203 Czech Republic, guarantor, belonging to the institution)
Edition
Analytical Chemistry, American Chemical Society, 2019, 0003-2700
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10406 Analytical chemistry
Country of publisher
United States of America
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 6.785
RIV identification code
RIV/00216224:14310/19:00111447
Organization unit
Faculty of Science
UT WoS
000495469100023
Keywords in English
MALDI MSI; LSCM; IHC; multimodal imaging; spheroids; coregistration; image analysis
Tags
International impact, Reviewed
Změněno: 16/4/2020 11:29, prof. Mgr. Jan Preisler, Ph.D.
Abstract
V originále
In this paper, we present an easy-to-follow procedure for the analysis of tissue sections from 3D cell cultures (spheroids) by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) and laser scanning confocal microscopy (LSCM). MALDI MSI was chosen to detect the distribution of the drug of interest, while fluorescence immunohistochemistry (IHC) followed by LSCM was used to localize the cells featuring specific markers of viability, proliferation, apoptosis and metastasis. The overlay of the mass spectrometry (MS) and IHC spheroid images, typically without any morphological features, required fiducial-based coregistration. The MALDI MSI protocol was optimized in terms of fiducial composition and antigen epitope preservation to allow MALDI MSI to be performed and directly followed by IHC analysis on exactly the same spheroid section. Once MS and IHC images were coregistered, the quantification of the MS and IHC signals was performed by an algorithm evaluating signal intensities along equidistant layers from the spheroid boundary to its center. This accurate colocalization of MS and IHC signals showed limited penetration of the clinically tested drug perifosine into spheroids during a 24 h period, revealing the fraction of proliferating and promigratory/proinvasive cells present in the perifosine-free areas, decrease of their abundance in the perifosine-positive regions and distinguishing between apoptosis resulting from hypoxia/nutrient deprivation and drug exposure.
Links
| EF16_013/0001775, research and development project |
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| LM2015062, research and development project |
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| LM2015091, research and development project |
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| LQ1601, research and development project |
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| MUNI/A/1018/2018, interní kód MU |
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| MUNI/G/0974/2016, interní kód MU |
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