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Related Experiment Video

Updated: Jun 13, 2026

Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
07:52

Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma

Published on: January 9, 2019

Optimized processing of fine-needle lymph node biopsies for automated immunostaining.

Adam D Aulbach1, Cheryl L Swenson, Matti Kiupel

  • 1MPI Research Inc., 54943 North Main Street, Mattawan, MI 49071-9399, USA. adam.aulbach@mpiresearch.com

Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
|May 11, 2010
PubMed
Summary

A new method uses visual assessment of cell suspension turbidity to ensure adequate cytologic samples for canine lymphoma immunophenotyping. This reliable technique streamlines processing and improves diagnostic accuracy for antigen detection.

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Area of Science:

  • Veterinary Pathology
  • Immunocytochemistry
  • Canine Lymphoma Diagnostics

Background:

  • Accurate immunophenotyping of canine lymphoma is crucial for diagnosis and treatment.
  • Current methods for assessing sample adequacy for immunocytochemistry can be time-consuming.
  • A need exists for a straightforward, reliable technique for pre-analysis evaluation of cytologic specimens.

Purpose of the Study:

  • To develop and validate a technique for postcollection processing and evaluation of cytologic specimens for antigen detection.
  • To assess the utility of visual estimation of cell suspension turbidity for determining sample adequacy for immunocytochemical staining.
  • To establish a reliable method for immunophenotyping canine lymphoma using automated immunostaining.

Main Methods:

  • Developed a technique using visual assessment of cell suspension turbidity and light microscopy of cytospin preparations.
  • Collected fine-needle lymph node aspirates from 81 dogs with lymphoma.
  • Processed samples using standardized media and automated immunostaining with CD79a and CD3e antibodies.

Main Results:

  • Visual assessment of turbidity reliably predicted adequate cell concentration for immunocytochemistry.
  • Identified 63% B cell, 23% T cell, 4% mixed T and B cell, and 4% non-T/non-B cell lymphomas.
  • Achieved 94% diagnostic yield, with nondiagnostic samples primarily from early in the study before turbidity guidelines were established.
  • Immunocytochemistry results showed complete agreement with available immunohistochemical correlates.

Conclusions:

  • Visual assessment of cell suspension turbidity is a straightforward and reliable method for evaluating sample adequacy for canine lymphoma immunophenotyping.
  • This technique simplifies postcollection processing and enhances the diagnostic utility of immunocytochemical staining.
  • The method promotes wider adoption of immunocytochemical techniques in veterinary diagnostics.