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@article{1583136,
author = {Machálková, Markéta and Pavlatovská, Barbora and Michálek, Jan and Pruška, Adam and Štěpka, Karel and Nečasová, Tereza and Radaszkiewicz, Katarzyna Anna and Kozubek, Michal and Šmarda, Jan and Preisler, Jan and Navrátilová, Jarmila},
article_number = {21},
doi = {http://dx.doi.org/10.1021/acs.analchem.9b02462},
keywords = {MALDI MSI; LSCM; IHC; multimodal imaging; spheroids; coregistration; image analysis},
language = {eng},
issn = {0003-2700},
journal = {Analytical Chemistry},
title = {Drug Penetration Analysis in 3D Cell Cultures Using Fiducial-Based Semiautomatic Coregistration of MALDI MSI and Immunofluorescence Images},
url = {https://pubs.acs.org/doi/10.1021/acs.analchem.9b02462},
volume = {91},
year = {2019}
}
TY - JOUR
ID - 1583136
AU - Machálková, Markéta - Pavlatovská, Barbora - Michálek, Jan - Pruška, Adam - Štěpka, Karel - Nečasová, Tereza - Radaszkiewicz, Katarzyna Anna - Kozubek, Michal - Šmarda, Jan - Preisler, Jan - Navrátilová, Jarmila
PY - 2019
TI - Drug Penetration Analysis in 3D Cell Cultures Using Fiducial-Based Semiautomatic Coregistration of MALDI MSI and Immunofluorescence Images
JF - Analytical Chemistry
VL - 91
IS - 21
SP - 13475-13484
EP - 13475-13484
PB - American Chemical Society
SN - 00032700
KW - MALDI MSI
KW - LSCM
KW - IHC
KW - multimodal imaging
KW - spheroids
KW - coregistration
KW - image analysis
UR - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02462
L2 - https://pubs.acs.org/doi/10.1021/acs.analchem.9b02462
N2 - 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.
ER -
MACHÁLKOVÁ, Markéta, Barbora PAVLATOVSKÁ, Jan MICHÁLEK, Adam PRUŠKA, Karel ŠTĚPKA, Tereza NEČASOVÁ, Katarzyna Anna RADASZKIEWICZ, Michal KOZUBEK, Jan ŠMARDA, Jan PREISLER a Jarmila NAVRÁTILOVÁ. Drug Penetration Analysis in 3D Cell Cultures Using Fiducial-Based Semiautomatic Coregistration of MALDI MSI and Immunofluorescence Images. \textit{Analytical Chemistry}. American Chemical Society, roč.~91, č.~21, s.~13475-13484. ISSN~0003-2700. doi:10.1021/acs.analchem.9b02462. 2019.
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