Package of ImageJ macros for quantification of cardiac histology.
Image requirements: Gomori silver staining for reticulum, standard image formats like BMP, GIF, JPEG, PNG, or TIFF.
This macro measures the size of cardiomyocytes (Group 1) in different parameters on slides stained with Gomori silver staining. The results are reported in an Excel spreadsheet on the desktop, which contains the individual results as well as an average value. An overlay image and color-coded instances will also be created. In addition, the size of the nuclei of the cardiomyocytes (Group 2) and the size of the capillaries (Group 3) are measured. Furthermore, the number of capillaries bordering each cardiomyocyte is measured (capillary contacts).
Image requirements: Immunofluorescence staining (WGA (Channel: Green), Nuclei (e.g. DAPI) (Channel: blue), Capillaries (e.g. CD31) (Channel: red)), standard image formats like BMP, GIF, JPEG, PNG, or TIFF.
This macro measures the size of cardiomyocytes (Group 1) in different parameters on slides stained with WGA+CD31+DAPI. The results are reported in an Excel spreadsheet on the desktop, which contains the individual results as well as an average value. An overlay image and color-coded instances will also be created. In addition, the size of the nuclei of the cardiomyocytes (Group 2) and the size of capillaries (Group 3) are measured. Furthermore, the number of capillaries bordering each cardiomyocyte is measured (capillary contacts).
Image requirements: Immunofluorescence staining (WGA (Channel: Green), Nuclei (e.g. DAPI) (Channel: blue), intramyocyte or/and intramyocyte-nuclei signal (e.g. pro-ANP) (Channel: red)), standard image formats like BMP, GIF, JPEG, PNG, or TIFF.
This macro measures the size of cardiomyocytes (Group 1) in different parameters on slides stained with WGA+CD31+intramyozyte and intramyocyte-nuclei signal. The results are reported in an Excel spreadsheet on the desktop, which contains the individual results as well as an average value. An overlay image and color-coded instances will also be created. In addition, the size of the nuclei of the cardiomyocytes (Group 2) are measured. Furthermore an intramyocyte and intramyocyte-nuclei signal in channel red is measured, the result is reported as %-value per instance.
- Open the image to be analyzed in ImageJ
- Start the Macro (also possible through shortcut "q")
- Find a suitable tolerance. (See instructions of the Pop-up window)
- Complete missing lines of segmentation, if necessary. Use "Freehand line" and add them to the roi manager. (See instructions of the Pop-up window)
- Summarized results and individual results are plotted in an Excel spreadsheet on the desktop. In addition, an overlay image with color-coded instances is created.
- If necessary, with the table and the overlayer image, incorrectly segmented instances can be excluded. Further calculation and analysis can then be done individually in the excel sheet.
For more information: HeartJ_Instructions.pdf
HearJ is based on a publication. If you use it successfully for your research, please cite us: https://doi.org/10.1186/s12967-023-04544-2
The publication contains further information and examples.