Leica MM COUNT NUCLEI CELL SCORING, MM COUNT NUCLEI, CELL SCORING User Manual

Leica MM Count Nuclei and
Cell Scoring powered by MetaMorph

Analysis Software Drop-in for Leica MM AF

• Ideal for two-wavelength experiments

• Count nuclei of most cell types

• Adaptive background correction™ for improved segmentation

• Cell-by-Cell measurements

Two popular and robust application modules are available together
in Leica MM AF to facilitate easy and fast segmentation analysis.

The Leica MM Count Nuclei Application Module for the Leica MM
AF is designed to automate accurate counting of nuclei of most
types of cells.

The module counts nuclei even when the background is uneven,
providing superior segmentation compared to simple threshold­ing.

The Leica MM Cell Scoring Module for the Leica MM AF is de­signed for the identication of two subpopulation of cells.

The module is an ideal tool for two-wavelength experiments. Using
a uorescent marker for all nuclei and an addition marker for cells
of interest, each wavelength is segmented and measured sepa­rately. The second stain can label the nucleus, cytoplasm or both.

The modules include Adaptive Background Correction, adapting
the cell detection algorithm to the local intensity ranges outside
the cells to provide the most robust segmentation available. This
technique enables cell detection even with highly variable back­ground uorescence within a single image.

A simple interface minimizes setup efforts and settings can be
saved for future use or customized to t your experiment.

®

Count nuclei analysis configuration

Select a sample image to congure your
settings

Count nuclei image analysis

Nuclei are identied, taking into consideration
nuclear size and varying background

Cell scoring acquisition

Acquisition of two wavelengths

Easy configuration for analysis

Count Nuclei setup is just two steps:

1. Select minimum and maximum nuclear size

2. Set the sample intensity level above the local
background

Cell Scoring setup involves a few easy steps:

1. Select the image of interest

2. Specify the size range of cells

3. Set the intensity above background

4. Choose the second image containing the
positive marker stain

5. Select the stain area (nucleus, cytoplasm

or both)

6. Specify the size range of cells and intensity
above background

7. Optionally congure the data log

Interactive data display

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

Multi-parameter analysis

Measurements for Count Nuclei include:

• Count of nuclei

• Integrated and average intensity

• Total and mean area

Measurements for Cell Scoring include:

• Count of negative and positive cells

• Percentage of negative and positive cells

• Total and mean nuclei area of negative and po­sitive cells

• Wavelength-specic integrated and average
intensity of negative and positive cells

Powerful data export

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

“MetaMorph® is a Registered Trademark of MDS

Analytical Technologies”

Cell scoring image analysis

The module identies the two different probes

www.leica-microsystems.com

Customization through journaling

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.