Standard Practice for Tissue Cryosection Analysis with SIMS

SCOPE
1.1 This practice provides the Secondary Ion Mass Spectrometry (SIMS) analyst with a method for analyzing tissue cryosections in the imaging mode of the instrument. This practice is suitable for frozen-freeze-dried and frozen-hydrated cryosection analysis.  
1.2 This practice does not describe methods for optimal freezing of the specimen for immobilizing diffusible chemical species in their native intracellular sites.  
1.3 This practice does not describe methods for obtaining cryosections from a frozen specimen.  
1.4 This practice is not suitable for any plastic embedded tissues.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

General Information

Status
Historical
Publication Date
09-May-1997
Technical Committee
Drafting Committee
Current Stage
Ref Project

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ASTM E1880-97 - Standard Practice for Tissue Cryosection Analysis with SIMS
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NOTICE: This standard has either been superseded and replaced by a new version or discontinued.
Contact ASTM International (www.astm.org) for the latest information.
Designation: E 1880 – 97
Standard Practice for
Tissue Cryosection Analysis with SIMS
This standard is issued under the fixed designation E 1880; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope top layer provides “fluffy” indium that helps in holding
cryosections flat for SIMS analysis.
1.1 This practice provides the Secondary Ion Mass Spec-
trometry (SIMS) analyst with a method for analyzing tissue
5. Significance and Use
cryosections in the imaging mode of the instrument. This
5.1 Pressing cryosections flat onto a conducting substrate
practice is suitable for frozen-freeze-dried and frozen-hydrated
has been one of the most challenging problems in SIMS
cryosection analysis.
analysis of cryogenically prepared tissue specimens. Frozen
1.2 This practice does not describe methods for optimal
cryosections often curl or peel off, or both, from the substrate
freezing of the specimen for immobilizing diffusible chemical
during freeze-drying. The curling of cryosections results in an
species in their native intracellular sites.
uneven sample surface for SIMS analysis. Furthermore, if
1.3 This practice does not describe methods for obtaining
freeze-dried cryosections are not attached tightly to the sub-
cryosections from a frozen specimen.
strate, the impact of the primary ion beam may result in further
1.4 This practice is not suitable for any plastic embedded
curling and even dislodging of the cryosection from the
tissues.
substrate. These problems render SIMS analysis difficult,
1.5 This standard does not purport to address all of the
frustrating and time consuming. The use of indium as a
safety concerns, if any, associated with its use. It is the
substrate for pressing cryosections flat has provided a practical
responsibility of the user of this standard to establish appro-
approach for analyzing cryogenically prepared tissue speci-
priate safety and health practices and determine the applica-
mens.
bility of regulatory limitations prior to use.
5.2 The procedure described herein has been successfully
2. Referenced Documents used for SIMS imaging of calcium transport in intestinal
,
4 5
tissue.
2.1 ASTM Standards:
5.3 The procedure described here is amenable to soft tissues
E 673 Terminology Related to Surface Analysis
of both animal and plant origin.
3. Terminology
6. Apparatus
3.1 Definitions:
6.1 The procedure described here can be used for tissue
3.1.1 See Terminology E 673 for definitions of terms used in
cryosection analysis with virtually any SIMS instrument.
SIMS.
6.2 A cold stage in the SIMS instrument is needed to
4. Summary of Practice
analyze frozen-hydrated specimens.
4.1 This practice describes a method for the analysis of
7. Procedure
tissue cryosections with SIMS. Tissue cryosections for SIMS
7.1 Prepare the indium substrate by pressing sheet indium
analysis need to be mounted flat on an electrically conducting
onto polished silicon wafer pieces of approximately 15 to 25
substrate. Cryosections should remain flat and adhere well to
mm surface area, which can be irregularly shaped. Next,
the substrate for SIMS analysis. This is achieved by pressing
frozen cryosections into an indium substrate. Indium, being a
malleable metal (Moh hardness 5 1.2, Young’s modu-
Sod, E. W., Crooker, A. R., and Morrison, G. H., “Biological Cryosection
lus 5 10.6 GPa), provides a “cushion” for pressing and holding
Preparation and Practical Ion Yield Evaluation for Ion Microscopic Analysis,”
the frozen cryosections flat for SIMS analysis. Indium sub- Journal of Microscopy (Oxford), Vol 160, 1990, p. 55.
Chandra, S., Fullmer, C. S., Smith, C. A., Wasserman, R. H., and Morrison, G.
strates are prepared by pressing sheet indium onto a polished
H. “Ion Microscopic Imaging of Calcium Transport in the Intestinal Tissue of
silicon wafer. An approximately 1 μm thick layer of indium
Vitamin D-deficient and Vitamin D-replete Chickens: A Ca Stable Isotope Study,”
(99.999 % purity) is then vapor deposited on this surface. This
Proceedings of the National Academy of Sciences (USA), Vol 87, 1990, p. 5715.
Chandra, S., and Morrison, G. H., “Sample Preparation of Animal Tissues and
Cell Cultures for Secondary Ion Mass Spectrometry (SIMS) Microscopy,” Biology
This practice is under the jurisdiction of ASTM Committee
...

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