A distinctive feature of the early stages of apoptosis is the disruption of the mitochondria, including changes in membrane and redox potential. Life Technologies offers a number of fluorescent probes for analyzing aspects of mitochondrial activity in live cells, including calcium flux, superoxide production, transition pore opening, and membrane potential.
Mitochondrial Apoptosis Assays
With minimal disruption of cellular function some assays can be monitored in real time while others are fixable for multiplexing or HCS applications.
|
 Detection of superoxide in live cells using MitoSOX™ Red superoxide indicator. |
|
MitoSOX™ Red Mitochondrial Superoxide Indicator, for live-cell imaging
|
Rhod-2 AM, cell permeant
|
Image-iT® LIVE Mitochondrial Transition Pore Assay Kit
|
|||
|---|---|---|---|---|---|
| Readout |
Targeted to mitochondria, exhibits fluorescent increase with superoxide concentration
|
Calcium indicator targeted to mitochondia; increases fluorescence with Ca
2+ concentration
|
Mitochondrial signal decreases with opening of transition pore
|
||
| Indicator |
MitoSOX™ Red
|
Rhod-2 AM
|
Hoechst 33342
|
Calcein
|
MitoTracker® Red CMXRos
|
| Common filter |
RFP/TRITC
|
TRITC
|
DAPI
|
FITC
|
TRITC
|
| Ex/Em (nm) |
510/580
|
552/577
|
350/461
|
494/517
|
579/599
|
| Photostability |
|
|
|
||
| Signal-to-noise ratio |
|
NA
|
|
||
| Bibliography |
Citations
|
Citations
|
Citations
|
||
| Fluorescence Microscope |
Yes
|
Yes
|
Yes
|
||
| HTS |
No
|
No
|
No
|
||
| Live cells |
Yes
|
Yes
|
Yes
|
||
| Fixable |
No
|
No
|
No
|
||
| Detergent resistant |
No
|
No
|
Yes
|
||
| Format |
10 x 50 µg
|
20 x 50 µg
|
1 kit (1,000 slides)
|
||
| Cat. No. |
M36008
|
R1245MP
|
I35103
|
||
| Readout |
Dye sequestered by active mitochondria in response to membrane potential
|
Mitochondrial depolarization causes JC-1 molecules to aggregate shifting from green to red emission
|
Mitotoxicity assay with orange stain for active mitochondria, cell-impermeant green stain for labeling dead cells, and a nuclear stain for localization
|
|||
| Indicator |
TMRM
|
JC-1
|
JC-1
|
Hoechst 33342
|
Image-iT® DEAD Green™ viability stain
|
MitoHealth stain
|
| Common filter |
TRITC
|
TRITC
|
TRITC
|
DAPI
|
FITC
|
TRITC
|
| Ex/Em (nm) |
552/576
|
514/529
|
514/590
|
350/461
|
488/515
|
550/580
|
| Photostability |
|
|
|
|||
| Signal-to-noise ratio |
NA
|
|
|
|||
| Bibliography |
Citations
|
Citations
|
||||
| Fluorescence microscope |
Yes
|
Yes
|
Yes
|
|||
| HCS |
Yes
|
|||||
| HTS |
No
|
No
|
No
|
|||
| Live cells |
Yes
|
Yes
|
Yes
|
|||
| Fixable |
No
|
No
|
Yes
|
|||
| Detergent resistant |
No
|
No
|
No
|
|||
| Protocol |
Microscopy
|
|||||
| Format |
25 mg
|
5 mg
|
2 x 96-well plates
|
|||
| Cat. No. |
T669
|
T3168
|
H10295
|
|||
| Readout |
Dye sequestered by active mitochondria in response to membrane potential
|
| Indicator |
MitoTracker® Orange
|
| Common filter |
TRITC
|
| Ex/Em (nm) |
550/580
|
| Photostability |
|
| Signal-to-noise ratio |
NA
|
| Multiplexing |
Yes
|
| Live cells |
Yes
|
| Fixed cells |
No
|
| Fixable |
Yes
|
| Detergent resistant |
No
|
| Protocol |
Microscopy
|
| Format |
20 x 50 μg
|
| Cat. No. |
M7510
|
|
MitoSOX™ Red Mitochondrial Superoxide Indicator, for live-cell imaging
|
Rhod-2 AM, cell permeant
|
Image-iT® LIVE Mitochondrial Transition Pore Assay Kit
|
|||
|---|---|---|---|---|---|
| Readout |
Targeted to mitochondria, exhibits fluorescent increase with superoxide concentration
|
Calcium indicator targeted to mitochondia; increases fluorescence with Ca
2+ concentration
|
Mitochondrial signal decreases with opening of transition pore
|
||
| Indicator |
MitoSOX™ Red
|
Rhod-2 AM
|
Hoechst 33342
|
Calcein
|
MitoTracker® Red CMXRos
|
| Common filter |
RFP/TRITC
|
TRITC
|
DAPI
|
FITC
|
TRITC
|
| Ex/Em (nm) |
510/580
|
552/577
|
350/461
|
494/517
|
579/599
|
| Photostability |
|
|
|
||
| Signal-to-noise ratio |
|
NA
|
|
||
| Bibliography |
Citations
|
Citations
|
Citations
|
||
| Fluorescence Microscope |
Yes
|
Yes
|
Yes
|
||
| HTS |
No
|
No
|
No
|
||
| Live cells |
Yes
|
Yes
|
Yes
|
||
| Fixable |
No
|
No
|
No
|
||
| Detergent resistant |
No
|
No
|
Yes
|
||
| Format |
10 x 50 µg
|
20 x 50 µg
|
1 kit (1,000 slides)
|
||
| Cat. No. |
M36008
|
R1245MP
|
I35103
|
||
| Readout |
Dye sequestered by active mitochondria in response to membrane potential
|
Mitochondrial depolarization causes JC-1 molecules to aggregate shifting from green to red emission
|
Mitotoxicity assay with orange stain for active mitochondria, cell-impermeant green stain for labeling dead cells, and a nuclear stain for localization
|
|||
| Indicator |
TMRM
|
JC-1
|
JC-1
|
Hoechst 33342
|
Image-iT® DEAD Green™ viability stain
|
MitoHealth stain
|
| Common filter |
TRITC
|
TRITC
|
TRITC
|
DAPI
|
FITC
|
TRITC
|
| Ex/Em (nm) |
552/576
|
514/529
|
514/590
|
350/461
|
488/515
|
550/580
|
| Photostability |
|
|
|
|||
| Signal-to-noise ratio |
NA
|
|
|
|||
| Bibliography |
Citations
|
Citations
|
||||
| Fluorescence microscope |
Yes
|
Yes
|
Yes
|
|||
| HCS |
Yes
|
|||||
| HTS |
No
|
No
|
No
|
|||
| Live cells |
Yes
|
Yes
|
Yes
|
|||
| Fixable |
No
|
No
|
Yes
|
|||
| Detergent resistant |
No
|
No
|
No
|
|||
| Protocol |
Microscopy
|
|||||
| Format |
25 mg
|
5 mg
|
2 x 96-well plates
|
|||
| Cat. No. |
T669
|
T3168
|
H10295
|
|||
| Readout |
Dye sequestered by active mitochondria in response to membrane potential
|
| Indicator |
MitoTracker® Orange
|
| Common filter |
TRITC
|
| Ex/Em (nm) |
550/580
|
| Photostability |
|
| Signal-to-noise ratio |
NA
|
| Multiplexing |
Yes
|
| Live cells |
Yes
|
| Fixed cells |
No
|
| Fixable |
Yes
|
| Detergent resistant |
No
|
| Protocol |
Microscopy
|
| Format |
20 x 50 μg
|
| Cat. No. |
M7510
|
Resources |
|
| Molecular Probes® Handbook BioProbes® Journal Articles | Related Products |