( ...)In Situ Cell Death Detection Kit

The In Situ Cell Death Detection Kit has been designed as a precise, fast, and simple non radioactive technique to detect and quantify apoptotic cell death at the single-cell level in tissues. It is based on the so-called TUNEL method.

During apoptosis, DNAse activity not only generates double-stranded, low-molecular-weight DNA fragments (mono- and oligonucleosomes), but also introduces strand breaks ("nicks") into the high-molecular-weight DNA. These processes can be identified by labeling the free 3-OH termini with terminal transferase (TdT), which attaches labeled nucleotides to all 3OH-ends (TUNEL reaction; TdT-mediated dUTP nick end labeling). This labeling is more sensitive than other methods, and is the method used by the Roche Applied Science In situ Cell Death Detection Kits. There are four different versions of this kit: In Situ Cell Death Detection Kit, Fluorescein; In Situ Cell Death Detection Kit, TMR red (tetramethylrhodamine red); In Situ Cell Death Detection Kit, AP (alkaline phosphatase); and In Situ Cell Death Detection Kit, POD (peroxidase).

This figure shows the detection principle of the TUNEL kits. Labeling with fluorescein may also be followed by immunohistochemical detection using anti-fluorescein-specific antibodies that are conjugated to POD or AP. Typical applications of the TUNEL kits are:

• Detection of individual apoptotic cells in frozen and formalin-fixed tissue sections in basic research and routine pathology
• Determination of the sensitivity of malignant cells to drug-induced apoptosis in cancer research and clinical oncology
• Typing of cells that are undergoing cell death (in heterogeneous populations) by double-staining procedures

Typical samples that can be examined with the In Situ Cell Death Detection Kits include:

• Cell smear preparations
• Adherent cells cultured on chamber slides
• Frozen, formalin-fixed, or paraffin-embedded tissue sections

The In Situ Cell Death Detection Kits, Fluorescein and TMR red, can also be used with cells in suspension (e.g., for analysis by flow cytometry).



This figure shows apoptotic cells in rabbit endometrium. The In Situ Cell Death Detection Kit, Fluorescein was used for staining a paraffin-embedded tissue section, which was then analyzed under a fluorescence microscope.


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