Civil Engineering Reference
In-Depth Information
Table 7.4 Examples of toxicity paradigms, possible analytes, readout modes and potential problems when using various readouts
for MNM toxicity screening
Toxicity type
or paradigm
Analyte
Probes
Readout mode
Utility
Potential problem
Cytotoxicity
Cell number/
proliferation
Hoechst 33342/
DAPI
Fluorimetry/
High content
assay
Cell quantifi cation,
nuclear content
Background signal from NPs
with blue fl uorescence
Membrane
leakage
Propidium
Iodide/Syto 9
Fluorimetry/
High content
assay
Compromised cell
membrane integrity
Background signal from NPs
with red fl uorescence (e.g.,
QDs)
Membrane
integrity
LDH assay
Absorbance at
490 nm
Cell viability
NPs may inhibit enzyme and/
or absorb at 490 nm
(CNTs, Ag)
ATPlite TM
ATP
Luminescence
Mitochondrial activity
and viability status
of cells
Not appropriate for NPs that
may inhibit enzyme and/or
absorb light (e.g., CNTs)
Mitochondrial
membrane
potential
JC1/TMRM/
chloromethyl-
X-rodhamine
Fluorimetry/
High content
assay
Loss of MMP
Background signal from NPs
with red or green
fl uorescence
Metabolic
activity
MTT, WST-1,
XTT.
Absorbance
Mitochondrial activity
and viability status
of cells
NPs may inhibit enzyme and/
or absorb light or
substrates
Intracellular
calcium fl ux
Fluo-4/ Fura
2-AM/Rho
2-AM
Fluorometry/
High content
assay
Increased Intra-
cellular calcium
level
Background signal from NPs
with green fl uorescence
Apoptosis
Calcein-AM
Fluorometry/
High content
assay
Mitochondrial
membrane
permeability
transition (MPT)
Background signal from NPs
with green fl uorescence
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