Molecular Imaging and Biology, том 20, издание 3, номера страниц: 368-377

Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay

Тип документаJournal Article
Дата публикации2017-12-21
ИздательSpringer Nature
Название журналаMolecular Imaging and Biology
Квартиль по SCImagoQ2
Квартиль по Web of ScienceQ2
Импакт-фактор 20213.48
ISSN15361632, 18602002
Cancer Research
Oncology
Radiology Nuclear Medicine and imaging
Краткое описание
PurposeHigh- and medium-throughput assays are now routine methods for drug screening and toxicology investigations on mammalian cells. However, a simple and cost-effective analysis of cytotoxicity that can be carried out with commonly used laboratory equipment is still required.ProceduresThe developed cytotoxicity assays are based on human cell lines stably expressing eGFP, tdTomato, mCherry, or Katushka2S fluorescent proteins. Red fluorescent proteins exhibit a higher signal-to-noise ratio, due to less interference by medium autofluorescence, in comparison to green fluorescent protein. Measurements have been performed on a fluorescence scanner, a plate fluorimeter, and a camera photodocumentation system.ResultsFor a 96-well plate assay, the sensitivity per well and the measurement duration were 250 cells and 15 min for the scanner, 500 cells and 2 min for the plate fluorimeter, and 1000 cells and less than 1 min for the camera detection. These sensitivities are similar to commonly used MTT (tetrazolium dye) assays. The used scanner and the camera had not been previously applied for cytotoxicity evaluation. An image processing scheme for the high-resolution scanner is proposed that significantly diminishes the number of control wells, even for a library containing fluorescent substances. The suggested cytotoxicity assay has been verified by measurements of the cytotoxicity of several well-known cytotoxic drugs and further applied to test a set of novel bacteriotoxic compounds in a medium-throughput format.ConclusionThe fluorescent signal of living cells is detected without disturbing them and adding any reagents, thus allowing to investigate time-dependent cytotoxicity effects on the same sample of cells. A fast, simple and cost-effective assay is suggested for cytotoxicity evaluation based on mammalian cells expressing fluorescent proteins and commonly used laboratory equipment.
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1. Kalinina M.A. и др. Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay // Molecular Imaging and Biology. 2017. Т. 20. № 3. С. 368–377.
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TY - JOUR

DO - 10.1007/s11307-017-1152-0

UR - http://dx.doi.org/10.1007/s11307-017-1152-0

TI - Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay

T2 - Molecular Imaging and Biology

AU - Kalinina, Marina A.

AU - Skvortsov, Dmitry A.

AU - Rubtsova, Maria P.

AU - Komarova, Ekaterina S.

AU - Dontsova, Olga A.

PY - 2017

DA - 2017/12/21

PB - Springer Science and Business Media LLC

SP - 368-377

IS - 3

VL - 20

SN - 1536-1632

SN - 1860-2002

ER -

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@article{2017,

doi = {10.1007/s11307-017-1152-0},

url = {https://doi.org/10.1007%2Fs11307-017-1152-0},

year = 2017,

month = {dec},

publisher = {Springer Science and Business Media {LLC}},

volume = {20},

number = {3},

pages = {368--377},

author = {Marina A. Kalinina and Dmitry A. Skvortsov and Maria P. Rubtsova and Ekaterina S. Komarova and Olga A. Dontsova},

title = {Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay}

}

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Kalinina, Marina A. et al. “Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay.” Molecular Imaging and Biology 20.3 (2017): 368–377. Crossref. Web.