MIBI staining v2

2021 ◽  
Author(s):  
Marc MB Bosse ◽  
Sean Bendall ◽  
Mike Angelo
Keyword(s):  
State Of The Art ◽  
Ion Beam ◽  
Time Of Flight ◽  
Ffpe Tissue ◽  
Staining Procedure ◽  
Glutaraldehyde Fixation ◽  
Output Data ◽  
Immunohistochemistry Staining ◽  
Imaging Time

This protocol is the standard FFPE tissue staining procedure recommended for Multiplex Ion Beam Imaging Time of Flight instrument (MIBI_TOF) developed in the Sean C. Bendall and Michael R. Angelo labs. The protocol has been successfully used for MIBI and is the result of extensive optimization experiments. It is inspired from state-of-the art of immunohistochemistry staining procedures but differs in some very important steps, namely, glutaraldehyde fixation and final washes prior tissue dehydration. Failure to follow exactly all steps described in this procedure may result in inconsistencies in output data after MIBI_TOF acquisition.

Exchange of Fluorinated Cyanine Dyes between Different Types of Silver Halide Microcrystals Studied by Imaging Time-of-Flight Secondary Ion Mass Spectrometry

Langmuir ◽  
2001 ◽  
Vol 17 (23) ◽  
pp. 7332-7338 ◽  
Author(s):  
Jens Lenaerts ◽  
Geert Verlinden ◽  
Luc Van Vaeck ◽  
Renaat Gijbels ◽  
Ingrid Geuens ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Secondary Ion Mass Spectrometry ◽  
Silver Halide ◽  
Time Of Flight ◽  
Cyanine Dyes ◽  
Imaging Time ◽  
Different Types ◽  
Ion Mass Spectrometry ◽  
Secondary Ion

Imaging of Membrane Lipids in Single Cells by Imprint-Imaging Time-of-Flight Secondary Ion Mass Spectrometry

2003 ◽  
Vol 75 (14) ◽  
pp. 3429-3434 ◽  
Author(s):  
Peter Sjövall ◽  
Jukka Lausmaa ◽  
Håkan Nygren ◽  
Lennart Carlsson ◽  
Per Malmberg
Keyword(s):  
Mass Spectrometry ◽  
Secondary Ion Mass Spectrometry ◽  
Membrane Lipids ◽  
Single Cells ◽  
Time Of Flight ◽  
Imaging Time ◽  
Ion Mass Spectrometry ◽  
Secondary Ion

New Instrumentation in Argonne's Hvem-Tandem Facility: Expanded Capability for in Situ Ion Beam Studies+

1995 ◽  
Vol 396 ◽  
Author(s):  
Charles W. Allen ◽  
Loren L. Funk ◽  
Edward A. Ryan
Keyword(s):  
Scientific Community ◽  
State Of The Art ◽  
Ion Irradiation ◽  
Ion Beam ◽  
Voltage Electron ◽  
User Facility ◽  
Order Of Magnitude ◽  
New Instrumentation ◽  
Better Than

AbstractDuring 1995, a state-of-the-art intermediate voltage electron microscope (IVEM) has been installed in the HVEM-Tandem Facility with in situ ion irradiation capabilities similar to those of the HVEM. A 300 kV Hitachi H-9000NAR has been interfaced to the two ion accelerators of the Facility, with a spatial resolution for imaging which is nearly an order of magnitude better than that for the 1.2 MV HVEM which dates from the early 1970s. The HVEM remains heavily utilized for electron- and ion irradiation-related materials studies, nevertheless, especially those for which less demanding microscopy is adequate. The capabilities and limitations of this IVEM and HVEM are compared. Both the HVEM and IVEM are part of the DOE funded User Facility and therefore are available to the scientific community for materials studies, free of charge for non-proprietary research.

scholarly journals Aberration-corrected focused ion beam for time-of-flight secondary neutral mass spectrometry

2019 ◽  
Vol 12 (8) ◽  
pp. 085005
Author(s):  
Kosuke Nagata ◽  
Ken-ichi Bajo ◽  
Satoru Itose ◽  
Miyuki Matsuya ◽  
Morio Ishihara ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Time Of Flight ◽  
Aberration Corrected

scholarly journals MIBI-TOF: A multiplexed imaging platform relates cellular phenotypes and tissue structure

2019 ◽  
Vol 5 (10) ◽  
pp. eaax5851 ◽  
Author(s):  
Leeat Keren ◽  
Marc Bosse ◽  
Steve Thompson ◽  
Tyler Risom ◽  
Kausalia Vijayaragavan ◽  
...  
Keyword(s):  
Single Molecule ◽  
Spatial Information ◽  
Dynamic Range ◽  
Ion Beam ◽  
Time Of Flight ◽  
Tissue Structure ◽  
Regional Variability ◽  
Flight Mass Spectrometry ◽  
Cellular Phenotypes ◽  
And Function

Understanding tissue structure and function requires tools that quantify the expression of multiple proteins while preserving spatial information. Here, we describe MIBI-TOF (multiplexed ion beam imaging by time of flight), an instrument that uses bright ion sources and orthogonal time-of-flight mass spectrometry to image metal-tagged antibodies at subcellular resolution in clinical tissue sections. We demonstrate quantitative, full periodic table coverage across a five-log dynamic range, imaging 36 labeled antibodies simultaneously with histochemical stains and endogenous elements. We image fields of view up to 800 μm × 800 μm at resolutions down to 260 nm with sensitivities approaching single-molecule detection. We leverage these properties to interrogate intrapatient heterogeneity in tumor organization in triple-negative breast cancer, revealing regional variability in tumor cell phenotypes in contrast to a structured immune response. Given its versatility and sample back-compatibility, MIBI-TOF is positioned to leverage existing annotated, archival tissue cohorts to explore emerging questions in cancer, immunology, and neurobiology.

scholarly journals Time-Of-Flight ERDA for Depth Profiling of Light Elements

2020 ◽  
Vol 4 (4) ◽  
pp. 40
Author(s):  
Keisuke Yasuda
Keyword(s):  
Ion Beam ◽  
Time Of Flight ◽  
Depth Profiling ◽  
Depth Resolution ◽  
Semiconductor Detectors ◽  
Light Elements ◽  
Elastic Recoil ◽  
Measurement Sensitivity ◽  
Elastic Recoil Detection ◽  
Detection Analysis

The time-of-flight elastic recoil detection analysis (TOF-ERDA) method is one of the ion beam analysis methods that is capable of analyzing light elements in a sample with excellent depth resolution. In this method, simultaneous measurements of recoil ion energy and time of flight are performed, and ion mass is evaluated. The energy of recoil ions is calculated from TOF, which gives better energy resolution than conventional Silicon semiconductor detectors (SSDs). TOF-ERDA is expected to be particularly applicable for the analysis of light elements in thin films. In this review, the principle of TOF-ERDA measurement and details of the measurement equipment along with the performance of the instrumentation, including depth resolution and measurement sensitivity, are described. Examples of TOF-ERDA analysis are presented with a focus on the results obtained from the measurement system developed by the author.

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