FLIVO™ In Vivo Apoptosis Kits
In Vivo Apoptosis Detection
ICT offers a growing range of novel tools for in vivo apoptosis detection. Use our fluorescent in vivo apoptosis probes, FLIVO™, to assess levels of apoptosis in live animals.
Above: Apoptotic ischemic hepatic cells fluoresce green with FAM- FLIVO; nuclei appear blue with Dapi staining. Courtesy of Drs. Raffaele Cursio and Pascal Colosetti (INSERM U895 C3M E2, Nice, France).
FLIVOTM in vivo Apoptosis Kits provide a simple yet accurate method to detect apoptosis in vivo. The non-cytotoxic FLIVO (FLuorescence in vIVO probes are injectable fluorescent inhibitors of caspase activity. Upon intravenous injection, FLIVO labels apoptotic cells green or red in just 30 minutes. The resulting green or red fluorescent signal within samples is a direct measure of apoptosis that occurred at the time the reagent was injected.
FLIVO may be used in animal models to monitor the efficacy of treatment or the effects of disease. It has been used successfully in murine models of cancer, rat brain studies, and avian brain studies, among others (see citations below).- FLIVO™ is non-toxic, cell-permeant, and crosses the blood-brain barrier.
- FAM- FLIVO has a green fluorescent label, carboxyfluorescein; ex/em: 488/520 nm
- SR- FLIVO has a red fluorescent label, sulforhodamine B; ex/em: 565/590-600 nm.
- Products are available in small kits (enough for 6 rats or mice) and large kits (enough for 24 rats or mice).
- COMING SOON: NIR Tracers for Non-Invasive Imaging of Apoptosis
More Information:
- Read more about FLIVO cancer applications
- Read more about FLIVO neuroscience applications
Learn more by visiting ICT's version 1.0 FLIVO webpage
The successful use of FLIVO has been cited in these recent publications (in reverse chronological order):
Riol-Blanco L, Delgado-Martín C, Sánchez-Sánchez N, Alonso-C LM, Gutiérrez-López MD, Del Hoyo GM, Navarro J, Sánchez-Madrid F, Cabañas C, Sánchez-Mateos P, Rodríguez-Fernández JL. Immunological synapse formation inhibits, via NF-kappaB and FOXO1, the apoptosis of dendritic cells. Nat Immunol. 2009 Jul;10(7):753-60. Epub 2009 Jun 7.
Delgado-Martín C, Riol-Blanco L, Alonso-C L M, Rodríguez-Fernández J L. A protocol to detect apoptotic dendritic cells in murine lymph nodes using multiphoton microscopy. Nature Protocols. DOI: 10.1038/nprot.2009.133. June 2009. http://www.natureprotocols.com/2009/06/10/a_protocol_to_detect_apoptotic.php
Cursio, R., Miele, C., Filippa, N., Colosetti, P., Auberger, P., Van Obberghen, E., and Gugenheim, J. Tyrosine phosphorylation of insulin receptor substrates during ischemia/reperfusion-induced apoptosis in rat liver. Langenbecks Arch Surg. 394, 123-131 (2009).
Cursio, R., Colosetti, P., Auberger, P., and Gugenheim, J. Liver Apoptosis Following Normothermic Ischemia-Reperfusion: In Vivo Evaluation of Caspase Activity by FLIVO Assay in Rats. Transplantation Proceedings. 40, 2038–2041 (2008).
Griffin, R.J. et al. Use of a Fluorescently Labeled Poly-Caspase Inhibitor for In Vivo Detection of Apoptosis Related to Vascular-Targeting Agent Arsenic Trioxide for Cancer Therapy. Technology in Cancer Research and Treatment. 6, 651-654 (2007). View article or Download PDF
Tsai, Y.C. et al. The ubiquitin ligase gp78 promotes sarcoma metastasis by targeting KAI1 for degradation. Nature Medicine. 13, 1504-1509 (2007).
COMING SOON: NIR Tracers for Non-Invasive Imaging of Apoptosis
These pictures were taken through a window chamber of one live FSaII murine fibrosarcoma tumor (in vivo). Before any treatment was administered, FAM-FLIVO™ was injected into the mouse to determine how many of the tumor cells were apoptotic: caspase-positive cells turn green, caspase-negative cells are dark. The first picture
(left) was taken 45 minutes after FAM-FLIVO™ was injected. Very few of the cells turned green, indicating that most of the tumor is living. The mouse was then injected with a drug (arsenic trioxide, ATO) for 3 hours. FAM-FLIVO™ was again injected into the mouse, and the second picture (right) was taken 45 minutes later. Within a few hours of treatment, Dr. Robert Griffin (formerly at the U of MN, now at the U of AR) was able to assess the efficacy of the drug using FAM-FLIVO™: this dose of ATO induced apoptosis in most of the tumor cells.
Sample protocol:
- Expose your animals to your experimental condition, and create positive and negative controls if possible.
- Reconstitute the reagent with 50mcL DMSO to form the stock concentrate (which can be frozen for future use).
- Dilute the injection buffer 1:10 with diH2O and sterilize by filtration.
- Add 550mcL 1X injection buffer to the reagent.
- Inject ~100mcL of diluted reagent into each mouse.
- Let circulate 30-45 minutes.
- Examine tissues under a fluorescent microscope, or sacrifice and excise cells.
- If desired, label cells with an additional stain, like 7AAD or labeled antibody.
- If desired, fix, embed, or freeze cells.
- Analyze cells with a fluorescent microscope, plate reader or flow cytometer.