Immunoguided Microdissection Techniques

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2011 Jul 16

PMID 21761293

Citations 8

Authors

Michael A Tangrea

Jeffrey C Hanson

Robert F Bonner

Thomas J Pohida

Jaime Rodriguez-Canales

Michael R Emmert-Buck

Affiliations

Soon will be listed here.

Abstract

Over the past 15 years, laser-based microdissection has improved the precision by which scientists can procure cells of interest from a heterogeneous tissue section. However, for studies that require a large amount of material (e.g., proteomics) or for cells that are scattered and difficult to identify by standard histological stains, an immunostain-based, automated approach becomes essential. In this chapter, we discuss the use of immunohistochemistry (IHC) and immunofluorescence (IF) to guide the microdissection process via manual and software-driven auto-dissection methods. Although technical challenges still exist with these innovative approaches, we present here methods and protocols to successfully perform immuno-based microdissection on commercially available laser dissection systems.

Citing Articles

Effect of Antigen Retrieval on Genomic DNA From Immunodissected Samples.

Johann D, Shin I, Roberge A, Laun S, Peterson E, Liu M J Histochem Cytochem. 2022; 70(9):643-658.

PMID: 36129255 PMC: 9527476. DOI: 10.1369/00221554221124163.

Analysis of neuroendocrine clones in NSCLCs using an immuno-guided laser-capture microdissection-based approach.

Baldelli E, Mandarano M, Bellezza G, Petricoin 3rd E, Pierobon M Cell Rep Methods. 2022; 2(8):100271.

PMID: 36046628 PMC: 9421534. DOI: 10.1016/j.crmeth.2022.100271.

Robust Acquisition of Spatial Transcriptional Programs in Tissues With Immunofluorescence-Guided Laser Capture Microdissection.

Zhang X, Hu C, Huang C, Wei Y, Li X, Hu M Front Cell Dev Biol. 2022; 10:853188.

PMID: 35399504 PMC: 8990165. DOI: 10.3389/fcell.2022.853188.

Computer-Aided Laser Dissection: A Microdissection Workflow Leveraging Image Analysis Tools.

Hipp J, Johann D, Chen Y, Madabhushi A, Monaco J, Cheng J J Pathol Inform. 2019; 9:45.

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Extensive rewiring of epithelial-stromal co-expression networks in breast cancer.

Oh E, Christensen S, Ghanta S, Jeong J, Bucur O, Glass B Genome Biol. 2015; 16:128.

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References

Mouledous L, Hunt S, Harcourt R, Harry J, Williams K, Gutstein H . Proteomic analysis of immunostained, laser-capture microdissected brain samples. Electrophoresis. 2003; 24(1-2):296-302. DOI: 10.1002/elps.200390026. View

Grover A, Tangrea M, Woodson K, Wallis B, Hanson J, Chuaqui R . Tumor-associated endothelial cells display GSTP1 and RARbeta2 promoter methylation in human prostate cancer. J Transl Med. 2006; 4:13. PMC: 1420331. DOI: 10.1186/1479-5876-4-13. View

Hanson J, Gillespie J, Grover A, Tangrea M, Chuaqui R, Emmert-Buck M . Gene promoter methylation in prostate tumor-associated stromal cells. J Natl Cancer Inst. 2006; 98(4):255-61. DOI: 10.1093/jnci/djj051. View

Fend F, Kremer M, Quintanilla-Martinez L . Laser capture microdissection: methodical aspects and applications with emphasis on immuno-laser capture microdissection. Pathobiology. 2001; 68(4-5):209-14. DOI: 10.1159/000055925. View

Tangrea M, Mukherjee S, Gao B, Markey S, Du Q, Armani M . Effect of immunohistochemistry on molecular analysis of tissue samples: implications for microdissection technologies. J Histochem Cytochem. 2011; 59(6):591-600. PMC: 3201191. DOI: 10.1369/0022155411404704. View

Huang S . Immunohistochemical demonstration of hepatitis B core and surface antigens in paraffin sections. Lab Invest. 1975; 33(1):88-95. View

Shi S, Key M, Kalra K . Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections. J Histochem Cytochem. 1991; 39(6):741-8. DOI: 10.1177/39.6.1709656. View

Fend F, Emmert-Buck M, Chuaqui R, Cole K, Lee J, Liotta L . Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis. Am J Pathol. 1999; 154(1):61-6. PMC: 1853427. DOI: 10.1016/S0002-9440(10)65251-0. View

Emmert-Buck M, Bonner R, Smith P, Chuaqui R, Zhuang Z, Goldstein S . Laser capture microdissection. Science. 1996; 274(5289):998-1001. DOI: 10.1126/science.274.5289.998. View

10.

Bonner R, Cole K, Pohida T, Chuaqui R, Goldstein S, Liotta L . Laser capture microdissection: molecular analysis of tissue. Science. 1997; 278(5342):1481,1483. DOI: 10.1126/science.278.5342.1481. View

11.

Battifora H, Kopinski M . The influence of protease digestion and duration of fixation on the immunostaining of keratins. A comparison of formalin and ethanol fixation. J Histochem Cytochem. 1986; 34(8):1095-100. DOI: 10.1177/34.8.2426335. View

12.

Fassunke J, Majores M, Ullmann C, Elger C, Schramm J, Wiestler O . In situ-RT and immunolaser microdissection for mRNA analysis of individual cells isolated from epilepsy-associated glioneuronal tumors. Lab Invest. 2004; 84(11):1520-5. DOI: 10.1038/labinvest.3700165. View

13.

Ramos-Vara J, Beissenherz M . Optimization of immunohistochemical methods using two different antigen retrieval methods on formalin-fixed paraffin-embedded tissues: experience with 63 markers. J Vet Diagn Invest. 2000; 12(4):307-11. DOI: 10.1177/104063870001200402. View

14.

Nakamura N, Ruebel K, Jin L, Qian X, Zhang H, Lloyd R . Laser capture microdissection for analysis of single cells. Methods Mol Med. 2007; 132:11-8. DOI: 10.1007/978-1-59745-298-4_2. View

15.

Ordonez N, Manning Jr J, Brooks T . Effect of trypsinization on the immunostaining of formalin-fixed, paraffin-embedded tissues. Am J Surg Pathol. 1988; 12(2):121-9. DOI: 10.1097/00000478-198802000-00005. View

16.

Miettinen M . Immunostaining of intermediate filament proteins in paraffin sections. Evaluation of optimal protease treatment to improve the immunoreactivity. Pathol Res Pract. 1989; 184(4):431-6. DOI: 10.1016/S0344-0338(89)80039-1. View

17.

Huang S, Minassian H, More J . Application of immunofluorescent staining on paraffin sections improved by trypsin digestion. Lab Invest. 1976; 35(4):383-90. View

18.

Anthony R, Urban Jr J, Alem F, Hamed H, Rozo C, Boucher J . Memory T(H)2 cells induce alternatively activated macrophages to mediate protection against nematode parasites. Nat Med. 2006; 12(8):955-60. PMC: 1955764. DOI: 10.1038/nm1451. View

19.

Pinkus G, OConnor E, ETHERIDGE C, Corson J . Optimal immunoreactivity of keratin proteins in formalin-fixed, paraffin-embedded tissue requires preliminary trypsinization. An immunoperoxidase study of various tumours using polyclonal and monoclonal antibodies. J Histochem Cytochem. 1985; 33(5):465-73. DOI: 10.1177/33.5.2580883. View

20.

Liu Y, Wu J, Liu S, Zhuang D, Wang Y, Shou X . Immuno-laser capture microdissection of frozen prolactioma sections to prepare proteomic samples. Colloids Surf B Biointerfaces. 2009; 71(2):187-93. DOI: 10.1016/j.colsurfb.2009.02.005. View