Literature DB >> 33822482

A Simple Method for Creating a High-Content Microscope for Imaging Multiplexed Tissue Microarrays.

Shabnam Abtahi1, Neal R Gliksman2, John F Heneghan1, Steven P Nilsen1, Jeremy L Muhlich3, Jay Copeland4, Emil Rozbicki5, Chris Allan5, Pradeep K Dudeja6,7, Jerrold R Turner1.   

Abstract

High-throughput, high-content imaging technologies and multiplex slide scanning have become widely used. Advantages of these approaches include the ability to archive digital copies of slides, review slides as teams using virtual microscopy software, and standardize analytical approaches. The cost and hardware and software inflexibility of dedicated slide scanning devices can, however, complicate implementation. Here, we describe a simple method that allows any microscope to be used for slide scanning. The only requirements are that the microscope be equipped with a motorized filter turret or wheels (for multi-channel fluorescence) and a motorized xyz stage. This example uses MetaMorph software, but the same principles can be used with any microscope control software that includes a few standard functions and allows programming of simple command routines, or journals. The series of journals that implement the method perform key functions, including assistance in defining an unlimited number of regions of interest (ROI) and imaging parameters. Fully automated acquisition is rapid, taking less than 3 hr to image fifty 2.5-mm ROIs in four channels. Following acquisition, images can be easily stitched and displayed using open-source or commercial image-processing and virtual microscope applications.
© 2021 Wiley Periodicals LLC. Basic Protocol 1: Hardware and software configuration Basic Protocol 2: Create a preliminary scan Basic Protocol 3: Select, save, and position ROIs Basic Protocol 4: Determine and set autofocus parameters Basic Protocol 5: Acquire tiled images Basic Protocol 6: Review the scans Basic Protocol 7: Reimage ROIs as needed Basic Protocol 8: Stitch, stack, and assemble images Basic Protocol 9: Repeat scanning for multiplex immunostaining or background subtraction. © 2021 Wiley Periodicals LLC.

Entities:  

Keywords:  automated image acquisition; digital pathology; fluorescence microscopy; slide scanning; tissue microarray

Mesh:

Year:  2021        PMID: 33822482      PMCID: PMC8103673          DOI: 10.1002/cpz1.68

Source DB:  PubMed          Journal:  Curr Protoc        ISSN: 2691-1299


  32 in total

1.  Constructing tissue microarrays for research use.

Authors:  Martina Storz Schweizer; Lela Schumacher; Mark A Rubin
Journal:  Curr Protoc Hum Genet       Date:  2004-02

2.  Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D).

Authors:  Weizhe Li; Ronald N Germain; Michael Y Gerner
Journal:  Proc Natl Acad Sci U S A       Date:  2017-08-14       Impact factor: 11.205

3.  Cyclic Immunofluorescence (CycIF), A Highly Multiplexed Method for Single-cell Imaging.

Authors:  Jia-Ren Lin; Mohammad Fallahi-Sichani; Jia-Yun Chen; Peter K Sorger
Journal:  Curr Protoc Chem Biol       Date:  2016-12-07

4.  Colonic epithelial hPepT1 expression occurs in inflammatory bowel disease: transport of bacterial peptides influences expression of MHC class 1 molecules.

Authors:  D Merlin; M Si-Tahar; S V Sitaraman; K Eastburn; I Williams; X Liu; M A Hediger; J L Madara
Journal:  Gastroenterology       Date:  2001-06       Impact factor: 22.682

Review 5.  Tissue microarray technology for high-throughput molecular profiling of cancer.

Authors:  O P Kallioniemi; U Wagner; J Kononen; G Sauter
Journal:  Hum Mol Genet       Date:  2001-04       Impact factor: 6.150

6.  Optimized multiplex immunofluorescence single-cell analysis reveals tuft cell heterogeneity.

Authors:  Eliot T McKinley; Yunxia Sui; Yousef Al-Kofahi; Bryan A Millis; Matthew J Tyska; Joseph T Roland; Alberto Santamaria-Pang; Christina L Ohland; Christian Jobin; Jeffrey L Franklin; Ken S Lau; Michael J Gerdes; Robert J Coffey
Journal:  JCI Insight       Date:  2017-06-02

7.  Multiple immunofluorescence labelling of formalin-fixed paraffin-embedded (FFPE) tissue.

Authors:  David Robertson; Kay Savage; Jorge S Reis-Filho; Clare M Isacke
Journal:  BMC Cell Biol       Date:  2008-03-19       Impact factor: 4.241

8.  Highly multiplexed imaging of single cells using a high-throughput cyclic immunofluorescence method.

Authors:  Jia-Ren Lin; Mohammad Fallahi-Sichani; Peter K Sorger
Journal:  Nat Commun       Date:  2015-09-24       Impact factor: 14.919

9.  QuPath: Open source software for digital pathology image analysis.

Authors:  Peter Bankhead; Maurice B Loughrey; José A Fernández; Yvonne Dombrowski; Darragh G McArt; Philip D Dunne; Stephen McQuaid; Ronan T Gray; Liam J Murray; Helen G Coleman; Jacqueline A James; Manuel Salto-Tellez; Peter W Hamilton
Journal:  Sci Rep       Date:  2017-12-04       Impact factor: 4.379

10.  Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections.

Authors:  Maddalena Maria Bolognesi; Marco Manzoni; Carla Rossana Scalia; Stefano Zannella; Francesca Maria Bosisio; Mario Faretta; Giorgio Cattoretti
Journal:  J Histochem Cytochem       Date:  2017-07-10       Impact factor: 2.479

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