Leica MM CELL CYCLE User Manual

Leica MM Cell Cycle powered by MetaMorph

Analysis Software Drop-in for Leica MM AF

• Quantitation of cell cycle stages

• Adaptive Background Correction™ for improved segmentation

• Field and Cell-by-Cell data logging

®

Cells have multiple checkpoints that can impede progression past
the stages of G0/G1, G2 or the midpoint of M during the cell cycle.
When checkpoints are active and cells are challenged by DNA
damage, hypoxia, metabolic changes or spindle disruption, normal
cells will arrest. One common property of cancerous cells is the
loss of those checkpoints. When cells lose checkpoint control and
are challenged, they often undergo apoptosis.

The Leica MM Cell Cycle for Leica MM AF software from Leica
Microsystems is designed for the quantitation of cell cycle stage
for cells labeled with a DNA stain. Additionally, a mitosis-specic
probe may be used to better identify M-phase cells and an apopto­sis-specic probe may be used to identify cells undergoing apop­tosis.

Images can be acquired using one, two or three different wave­lengths for the DNA stain, M-phase marker and apoptosis marker.

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.

A 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.

Multiple wavelength acquisition

The software acquires multiple wavelengths
and color combines the images during visual­ization (right).

Robust analysis

Cell cycle classification can be performed with
a single or multi wavelength assay. Top: CHO-K1
cells stained with Hoechst 33342. Bottom: The
Cell Cycle module identifies cell cycle phases:
G0/G1 (dark blue), S (light blue), G2 (green),
Early M (orange) and Late M (red).

Configuration for analysis

Step 1. Select the DNA stain image.
Step 2. Specify the size range of cells and in-

tensity above local background.

Step 3. Set the classication criterion for mi-

tosis by specifying the intensity of the
DNA stain or an optional mitotic-speci­c stain.

Step 4. If using an apoptotic stain, select the

stain area, size range of cells and inten­sity above local background.

Step 5. Preview classication results and inter-

actively adjust cutoff values.

Step 6. Optionally set 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.

Customization through journaling

Multi-parameter analysis

The Application Module can generate a number
of eld or cell-by-cell parameters, including:

• Count and percentage of G0/G1, S, G2, early M,
late M and apoptotic cells

• DNA area, mitotic and apoptotic integrated and
average intensities

“MetaMorph® is a Registered Trademark of MDS

Analytical Technologies”

Interactive graphs

Cell classifications can be set by interactively
moving cutoff values directly on the graphs.

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.

Powerful data export capabilities

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

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