ASTM E1880-12(2020)

This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.
Designation: E1880 − 12 (Reapproved 2020)
Standard Practice for
Tissue Cryosection Analysis with SIMS
This standard is issued under the fixed designation E1880; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope 3.1.1 SeeTerminology E673 for definitions of terms used in
SIMS.
1.1 This practice provides the Secondary Ion Mass Spec-
trometry (SIMS) analyst with a method for analyzing tissue
4. Summary of Practice
cryosections in the imaging mode of the instrument. This
4.1 This practice describes a method for the analysis of
practice is suitable for frozen-freeze-dried and frozen-hydrated
tissue cryosections with SIMS. Tissue cryosections for SIMS
cryosection analysis.
analysis need to be mounted flat on an electrically conducting
1.2 This practice does not describe methods for optimal
substrate. Cryosections should remain flat and adhere well to
freezing of the specimen for immobilizing diffusible chemical
the substrate for SIMS analysis. This is achieved by pressing
species in their native intracellular sites.
frozen cryosections into an indium substrate. Indium, being a
1.3 This practice does not describe methods for obtaining
malleable metal (Moh hardness = 1.2,Young’s modulus = 10.6
cryosections from a frozen specimen.
GPa), provides a “cushion” for pressing and holding the frozen
cryosections flat for SIMS analysis. Indium substrates are
1.4 This practice is not suitable for any plastic embedded
prepared by pressing sheet indium onto a polished silicon
tissues.
wafer.An approximately 1 µm thick layer of indium (99.999 %
1.5 This standard does not purport to address all of the
purity) is then vapor deposited on this surface. This top layer
safety concerns, if any, associated with its use. It is the
provides “fluffy” indium that helps in holding cryosections flat
responsibility of the user of this standard to establish appro-
for SIMS analysis.
priate safety, health, and environmental practices and deter-
mine the applicability of regulatory limitations prior to use.
5. Significance and Use
1.6 This international standard was developed in accor-
5.1 Pressing cryosections flat onto a conducting substrate
dance with internationally recognized principles on standard-
has been one of the most challenging problems in SIMS
ization established in the Decision on Principles for the
analysis of cryogenically prepared tissue specimens. Frozen
Development of International Standards, Guides and Recom-
cryosections often curl or peel off, or both, from the substrate
mendations issued by the World Trade Organization Technical
during freeze-drying. The curling of cryosections results in an
Barriers to Trade (TBT) Committee.
uneven sample surface for SIMS analysis. Furthermore, if
freeze-dried cryosections are not attached tightly to the
2. Referenced Documents
substrate, the impact of the primary ion beam may result in
2.1 ASTM Standards:
further curling and even dislodging of the cryosection from the
E673 Terminology Relating to SurfaceAnalysis (Withdrawn
substrate. These problems render SIMS analysis difficult,
2012)
frustrating and time consuming. The use of indium as a
substrate for pressing cryosections flat has provided a practical
3. Terminology
approach for analyzing cryogenically prepared tissue speci-
3.1 Definitions:
mens (1).
5.2 The procedure described herein has been successfully
This practice is under the jurisdiction of ASTM Committee E42 on Surface
used for SIMS imaging of calcium and magnesium transport
Analysis and is the direct responsibility of Subcommittee E42.06 on SIMS.
and localization of anticancer drugs in animal models (2, 3, 4,
Current edition approved Dec. 1, 2020. Published December 2020. Originally
5).
approved in 1997. Last previous edition approved in 2012 as E1880 – 12. DOI:
10.1520/E1880-12R20.
5.3 The procedure described here is amenable to soft tissues
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
of both animal and plant origin.
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
3 4
The last approved version of this historical standard is referenced on The boldface numbers in parentheses refer to a list of references at the end of
www.astm.org. this standard.
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E1880 − 12 (2020)
6. Apparatus polished surface of the top silicon piece is used to press the
cryosection onto indium substrate in order to avoid introducing
6.1 The procedure described here can be used for tissue
the irregular topography of the rough silicon surface. Remove
cryosection analysis with virtually any SIMS instrument.
the top silicon piece by sliding it off using chilled tweezers.
6.2 A cold stage in the SIMS instrument is needed to
The pressed frozen cryosection on the indium substrate is now
analyze frozen-hydrated specimens (6).
ready for frozen-hydrated analysis with a cold stage in the
SIMS instrument.Alternatively, the pressed cryosection on the
7. Procedure
indium substrate can be freeze-dried by
...