Leica MM GRANULARITY User Manual

Leica MM Granularity powered by MetaMorph

Analysis Software Drop-in for Leica MM AF

• Counts nuclei and punctate

• Adaptive Background Correction™ for improved cell quantitation

• Choice of six granularity indices

• Field and Cell-by-Cell data logging

®

The Granularity Application Module for the Leica MM AF software
from Leica Microsystems is designed to facilitate the segmentation
and quantitation of punctate. This module can be used to analyze
images from common experiments such as studying intracellular
structures, receptor internalization or clustering target molecules.

The user can select from six granularity indices, choosing the one
that best suits the cell and receptor type, cell density and other
experiment parameters. The granule count, area and intensity per­cell as per-image can be captured.

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 granule 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.

Adaptive Background CorrectionTM for im­proved segmentation

Using standard thresholding can result in incor­rect grain counts (orange), Adaptive Background
Correction improves grain quantitation even in
cells with high background (red arrow).Assay
kit: Molecular Probes, Vybrant Assay Kit #7.

Pits (white) and nuclei (green) can be simultane­ously segmented, saving analysis time. Grains
are accurately identified even in cells with high
background (red arrow).

Configuration for analysis

Step 1. Select the granule image.
Step 2. Specify the size range of punctate and

intensity above local background.

Step 3. If using a nuclear stain, select the

nuclear image, size range of nuclei and
intensity above local background.

Step 4. 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

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

• Granule and Nuclear Count

• Granule and Nuclear Total and Mean Area

• Granule and Nuclear Integrated and Average
Intensity

The module can determine and log the following
six granularity indexes:

• Texture Index

• Cellular Texture Index

• Gradient Index

• Cellular Gradient Index

• Laplacian Index

• Cellular Laplacian Index

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
Analytical Technologies”

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