Leica MM MITOTIC INDEX User Manual

Leica MM Mitotic Index powered by MetaMorph

Analysis Software Drop-in for Leica MM AF

• Quantitation of mitotic and interphase cells

• Multi-parameter analysis of two probes

• Adaptive Background Correction™ for improved segmentation

• Cell-by-Cell measurements and data logging
Cells are labeled with a DNA stain and a uorescent mitosis-spe-

Cell-based assays for cell cycle status have become critical for
oncology research. For example, potential anti-cancer therapeu­tics rely on arresting mitosis in cancerous cells to prevent uncon­trolled proliferation.

In some forms of cancer, progression through mitosis can be
stopped by merely disrupting the mitotic spindle. In others, the
feedback is missing and cells proceed through mitosis without
proper chromosome segregation and accumulate mutations. Cell­based assays provide an efcient method for modeling the effects
of drugs on the cell cycle and quantitating the accumulation of
cells in mitosis. The Leica MM Mitotic Index module for the Leica
MM AF software is designed for the quantitative discrimination of
mitotic and interphase cells.

cic marker, such as immunouorescence staining for Histone 3
S10 phosphorylation. Two different wavelengths are acquired and
the images are analyzed with the Leica MM Mitotic module.

The module utilizes Adaptive Background Correction adapting the
cell detection algorithm to the local intensity ranges between and
within cells to provide the most robust segmentation available.
This technique enables probe detection even with highly variable
background uorescence within a single image.

The simple interface minimizes setup efforts and at the same time
enables users to customize the settings and measurements to ob­tain the best possible results specic to the type of cells used in
the experiments.

®

Easy Configuration for analysis

Customization through journaling

Multiple probe acquisition

CHO-K1 cells treated with Nocodazole for 18
hours before staining with anti-phospho-His­tone H3 (Ser28). Top: control, bottom: 50 ng/ml
Nocodazole.

Image analysis

The Leica MM Mitotic Index module identifies
mitotic cells (green) and interphase cells (red).

1. Select the nuclear stain image

2. Specify the cell size range

3. Set the intensity above local background

4. Select the mitotic nuclear stain image

5. Set the intensity above local background

6. Optionally choose the reporting parameters

Interactive data display

Once the analysis is run, the Cellular Results
table allows you to interactively view individual
cells’ data. Clicking one or multiple cells in the
image highlights the data for the selected cell(s)
in the table.

Journals are sophisticated and powerful macros
that record and perform a series of tasks without
the need for a programming language. The mod­ules can be incorporated into a Leica MM AF
journal to increase the customization and auto­mation of your analysis.

Multi-parameter analysis

The application module generates a number of
eld or cell-by-cell parameters, including:

• Count of mitotic and non-mitotic nuclei

• Percentage of mitotic and non-mitotic nuclei

• Total and mean nuclei area of mitotic and non­mitotic cells

• Wavelength-specic integrated and average
intensities of mitotic and non-mitotic nuclei

Powerful data export capabilities

All measurements can be directly exported to a
text le or Microsoft® Excel® for further analy­sis.

“MetaMorph® is a Registered Trademark of MDS Analyti-

cal Technologies”

Export data for further analysis

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