Advertisement

Image Analysis Algorithms for Immunohistochemical Assessment of Cell Death

  • Stan Krajewski
  • Jeffrey Wang
  • Tashmia Khan
  • Jonathan Liu
  • Chia-Hung Sze
  • Maryla Krajewska
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1254)

Abstract

Light microscopy allows for the inexpensive and fast detection of neuronal /glial cell demise and estimation of infarct and traumatic lesion volumes ; the direct correlates of cell death . Quantitative assessment of brain tissue damage following stroke , traumatic brain injury (TBI ) or neurodegenerative diseases , and recovery after therapeutic intervention has been facilitated by recent developments in computer-assisted image analysis technologies that enable more objective and accurate morphometric quantification of cell injury in whole brain sections . In this chapter, the proposed workflow describes what tasks need to be fulfilled to visualize and gauge cell death characterization by histological stains and immunohistochemical markers .

Key words

Artifact-free brainfixation/processing into cryo- or paraffin blocks Digital pathology TBI neuropathology and hemorrhagescores IHC Algorithm-based morphometry of death events 

Notes

Acknowledgements

This work was supported by the National Institutes of Health (NIH) Grant NS-036821 and the Department of Defense (DoD) Grant W81XWH-10-1-0847 awarded to Stan Krajewski. We are greatly indebted to Dr. Allen Olsen from Aperio Technology, Inc. for collaborative development and beta-testing of Cell/Cytoplasmic v2 algorithm. We thank Xianshu Huang for high-quality histological preparations and immunocytochemistry, Tomi Omel and Francisco Beltran for excellent animal care assistance.

References

  1. 1.
    Galluzzi L, Maiuri MC, Vitale I et al (2007) Cell death modalities: classification and pathophysiological implications. Cell Death Differ 14:1237–1243PubMedCrossRefGoogle Scholar
  2. 2.
    Vanden Berghe T, Grootjans S, Goossens V et al (2013) Determination of apoptotic and necrotic cell death in vitro and in vivo. Methods 61:117–129PubMedCrossRefGoogle Scholar
  3. 3.
    Kroemer G, Galluzzi L, Vandenabeele P et al (2009) Classification of cell death: recommendations of the Nomenclature Committee on Cell Death 2009. Cell Death Differ 16:3–11PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Rizzardi AE, Johnson AT, Vogel RI et al (2012) Quantitative comparison of immunohistochemical staining measured by digital image analysis versus pathologist visual scoring. Diagn Pathol 7:42PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Krajewska M, Smith LH, Rong J et al (2009) Image analysis algorithms for immunohistochemical assessment of cell death events and fibrosis in tissue sections. J Histochem Cytochem 57:649–663PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Weinstein RS (2009) Risks and rewards of pathology innovation: the academic pathology department as a business incubator. Arch Pathol Lab Med 133:580–586PubMedGoogle Scholar
  7. 7.
    Krajewska M, You Z, Rong J et al (2011) Neuronal deletion of caspase 8 protects against brain injury in mouse models of controlled cortical impact and kainic acid-induced excitotoxicity. PLoS One 6:e24341PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Brazdziute E, Laurinavicius A (2011) Digital pathology evaluation of complement C4d component deposition in the kidney allograft biopsies is a useful tool to improve reproducibility of the scoring. Diagn Pathol 6 (Suppl 1):S5PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Krajewska M, Xu L, Xu W et al (2011) Endoplasmic reticulum protein BI-1 modulates unfolded protein response signaling and protects against stroke and traumatic brain injury. Brain Res 1370:227–237PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Okuno S, Saito A, Hayashi T et al (2004) The c-Jun N-terminal protein kinase signaling pathway mediates Bax activation and subsequent neuronal apoptosis through interaction with Bim after transient focal cerebral ischemia. J Neurosci 24:7879–7887PubMedCrossRefGoogle Scholar
  11. 11.
    Silva RM, Ries V, Oo TF et al (2005) CHOP/GADD153 is a mediator of apoptotic death in substantia nigra dopamine neurons in an in vivo neurotoxin model of parkinsonism. J Neurochem 95:974–986PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Szegezdi E, Logue SE, Gorman AM, Samali A et al (2006) Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 7: 880–885PubMedCentralPubMedCrossRefGoogle Scholar
  13. 13.
    Yuan J (2006) Divergence from a dedicated cellular suicide mechanism: exploring the evolution of cell death. Mol Cell 23:1–12PubMedCrossRefGoogle Scholar
  14. 14.
    Blennow K, Hardy J, Zetterberg H (2012) The neuropathology and neurobiology of traumatic brain injury. Neuron 76:886–899PubMedCrossRefGoogle Scholar
  15. 15.
    Krajewska M, Rosenthal RE, Mikolajczyk J et al (2004) Early processing of Bid and caspase-6, -8, -10, -14 in the canine brain during cardiac arrest and resuscitation. Exp Neurol 189:261–279PubMedCrossRefGoogle Scholar
  16. 16.
    Ruifrok AC, Johnston DA (2001) Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol 23: 291–299PubMedGoogle Scholar
  17. 17.
    Yu J, Wang Z, Kinzler KW et al (2003) PUMA mediates the apoptotic response to p53 in colorectal cancer cells. Proc Natl Acad Sci U S A 100:1931–1936PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Paxinos G, Franklin K (2001) The mouse brain in stereotaxic coordinates. Academic, San DiegoGoogle Scholar
  19. 19.
    Rosen GD, Harry JD (1990) Brain volume estimation from serial section measurements: a comparison of methodologies. J Neurosci Methods 35:115–124PubMedCrossRefGoogle Scholar
  20. 20.
    Prieto DA, Hood BL, Darfler MM et al (2005) Liquid Tissue: proteomic profiling of formalin-fixed tissues. Biotechniques Suppl: 32–35Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Stan Krajewski
    • 1
    • 2
  • Jeffrey Wang
    • 1
  • Tashmia Khan
    • 1
  • Jonathan Liu
    • 1
  • Chia-Hung Sze
    • 1
  • Maryla Krajewska
    • 1
    • 2
  1. 1.Sanford-Burnham Medical Research InstituteLa JollaUSA
  2. 2.Cellestan-ImmunoQuant, Inc.OceansideUSA

Personalised recommendations