TABLE 1.
Single Fluorescent Markers Used to Study Human Malarias by Flow Cytometry
| Stain | Interacts with |
Excitation Maximum |
Emission Maximum |
Dose1 | Fixative | References |
|---|---|---|---|---|---|---|
|
Hoechst
33258 |
DNA | 345 | 478 | 2 μM, | 0.25% glutaraldehyde | (van Vianen et al., 1990)2 |
| 200 μM, | None | (Brown et al., 1980) | ||||
| 32.05 μM, | None | (Izumiyama et al., 2009) | ||||
|
| ||||||
|
Hoechst
33342 |
DNA | 355 | 465 | 4.06 μM | 0.25 % glutaraldehyde | (Dent et al., 2008)3 |
| 32.47 μM | None | (Izumiyama et al., 2009) | ||||
|
| ||||||
|
Green
Fluorescent Protein |
Varies | 3954 | 508 | NA | None | (Sanchez et al., 2007)2 |
|
| ||||||
| SYTO-9 | DNA/RNA | 485/486 | 598/501 | 2.5 μM | None | (Izumiyama et al., 2009) |
|
| ||||||
| SYBR Green I | DNA | 488 | 522 | 2X | 1% paraformaldehyde | (Izumiyama et al., 2009) |
| 10X 5 | 0.25% glutaraldehyde | (Dent et al., 2008)3 | ||||
|
| ||||||
| SYTO 16 | DNA/RNA | 488/494 | 518/525 | 100 Nm | 2% paraformaldehyde | (Dahl and Rosenthal, 2007) |
| 5 μM | 0.25% glutaraldehyde | (Jimenez-Diaz et al., 2009) | ||||
|
| ||||||
| YOYO-1 | Nucleic acids, with high affinity for DNA |
491 | 509 | 1 nM | 2% paraformaldehyde + 0.1% Triton X-100 |
(Dahl and Rosenthal, 2007)2 |
| 393.5 nM | 0.04% glutaraldehyde + 0.5 mg/ml RNAse |
(Li et al., 2007) | ||||
| 0.5 μM | 0.25% glutaraldehyde | (Angulo-Barturen et al., 2008) | ||||
|
| ||||||
|
Acridine
Orange |
DNA/RNA | 503 | 530/640 | 72.89 μM, | 1% paraformaldehyde + acid buffers |
(Hare, 1986) |
| 9.94 μM | None | (Izumiyama et al., 2009) | ||||
|
| ||||||
| SYTOX Green | Nucleic Acids |
504 | 523 | 5 μM | 4% formaldehyde + 0.0075% glutaraldehyde then 0.25% Triton X-100 |
(Chandramohanadas et al., 2009) |
|
| ||||||
|
Thiazole
Orange |
Primarily RNA |
509 | 533 | 100 nM | None | (Makler et al., 1987) |
|
| ||||||
|
Ethidium
Bromide |
Nucleic acids |
510 6 | 595 | 253.61 μM | None | (Staalsoe et al., 1999) |
| 25.4 μM | (Dent et al., 2008)3 | |||||
| 2.03 μM | (Persson et al., 2006)3 | |||||
|
| ||||||
|
Propidium
Iodide |
Nucleic Acids |
535 | 617 | 14.96 μM | 0.25% glutaraldehyde | (Pattanapanyasat et al., 1997)2 |
|
| ||||||
| Hydroethidine | Live/ DNA |
365/ 535 |
435/ 610 |
158.49 μM | None | (van der Heyde et al., 1995) |
|
| ||||||
| SYTO-61 | Nucleic Acids |
628 | 645 | 1μM | None | (Fu et al., 2010) |
1
Published concentrations have been converted to molar units
2
Used to determine IC50/90 of antimalarial compounds
3
Used for growth or invasion inhibition assays
4
GFP also has a 1/3 maximum excitation peak at 489 nm
5
SYBR Green is available as a 10,000 X stock from Invitrogen. The concentration is proprietary
6
Ethidium bromide has an additional excitation maximum at 302 nm which has yet to be employed for malaria cytometric methods