Biomedical Engineering Reference
In-Depth Information
Cell
seeding
Test
compound
addition
Assay
reagent
addition
Data
recording
Equilibration
Exposure
Assessment
10min-24hours
0-24 hours
1 hour-7days
Figure 10.1
Time-scale representation for toxicity assessment.
Table 10.1
Colorimetric assays for semi-quantitative analysis of cytotoxicity
Reagent
Process by live cells
Dissolved in
Wavelength (nm)
MTT
Conversion of yellow water soluble
tetrazolium salt to blue formazan
DMSO or isopropanol
570
XTT
Conversion of yellowish water
soluble tetrazolium salt to orange
formazan
Water or PBS
450-500
Crystal violet
Uptake of violet dye in less than
10 min by live cells
Acetic acid
570
Neutral red
Uptake of red dye and conversion
in lysosome
Acetic acid-ethanol
540
Alamar blue
Conversion of nonfluorescence
blue dye to red fluorescence
Water or PBS
Excitation: 530
Emission: 590
DMSO, dimethyl sulfoxide; MTT, 3-[4, 5-dimethyl thiazol-2-y1]-2, 5-diphenyl tetrazolium bromide; PBS,
phosphate buffer solution; XTT, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide.
Fluorescence imaging
Confocal microscopy
Optical microscopy
Electron microscopy
Cell
morphology
ISNT assay
Comet assay
TUNNEL assay
Cell cycle
(Flowcytometry)
DNA synthesis
(Thymidine, brdU)
Gene expression
(c-Myc, p53, Bcl-2,
caspase-3, ...)
LDH
FDA/PI
Crystal violet
Tr ypan blue
Hoechst
AO/PI
In vitro
cytotoxicity
assays
Genetic
indices
Membrance
integrity
Organelle
function
ROS determination
Lysosome (Neutral red)
Intracellular esterases (Calcein AM)
Mitochondria
(MTT, XTT, MTS, Alamar-blue)
Figure 10.2
In vitro
cell toxicity/viability assays in four categories: genetic indices, membrane
integrity, organelle function and cell morphology.
