148 lines
21 KiB
XML
148 lines
21 KiB
XML
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<mods:title id="6FF610655CA72F447141297287119F9F">Integrative microscopy to explore physical and nanomechanical properties of eggshells of diapausing embryos in Rotifera: a proof-of-concept study</mods:title>
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<mods:namePart id="7C003F1C961F2AB7E8EECED10A09787F">Araujo, Thiago Q.</mods:namePart>
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<mods:namePart id="569610ECF0718B241130D9F416B198DC">Walsh, Elizabeth J.</mods:namePart>
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<mods:namePart id="33734A9F26C74E7D52E504C4A94FD947">Wallace, Robert L.</mods:namePart>
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<treatment id="03B687E6220BFFE9FF42FB10FE7F7355" LSID="urn:lsid:plazi:treatment:03B687E6220BFFE9FF42FB10FE7F7355" httpUri="http://treatment.plazi.org/id/03B687E6220BFFE9FF42FB10FE7F7355" lastPageId="9" lastPageNumber="1993" pageId="7" pageNumber="1991">
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<heading id="D0E8819C220BFFE7FF42FB10FD88747B" bold="true" box="[160,626,1265,1291]" centered="true" fontSize="11" level="2" pageId="7" pageNumber="1991" reason="3">
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<emphasis id="B96BEAE2220BFFE7FF42FB10FD88747B" bold="true" box="[160,626,1265,1291]" pageId="7" pageNumber="1991">
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<subSubSection id="C305657B220BFFE7FF42FB10FE97747B" box="[160,365,1265,1291]" pageId="7" pageNumber="1991" type="discussion">
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<paragraph id="8BA036F0220BFFE7FF42FB10FE97747B" blockId="7.[160,626,1265,1291]" box="[160,365,1265,1291]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FB10FE9C747B" bold="true" box="[160,358,1265,1291]" italics="true" pageId="7" pageNumber="1991">
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Phylum
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<taxonomicName id="4C1F4D73220BFFE7FEE6FB10FE9C747B" ID-CoL="5Y" ID-ENA="10190" box="[260,358,1265,1291]" kingdom="Animalia" pageId="7" pageNumber="1991" phylum="Rotifera" rank="phylum">Rotifera</taxonomicName>
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</emphasis>
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:
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</paragraph>
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</subSubSection>
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<subSubSection id="C305657B220BFFE7FE97FB10FD88747B" box="[373,626,1265,1291]" pageId="7" pageNumber="1991" type="nomenclature">
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<paragraph id="8BA036F0220BFFE7FE97FB10FD88747B" blockId="7.[160,626,1265,1291]" box="[373,626,1265,1291]" pageId="7" pageNumber="1991">
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<taxonomicName id="4C1F4D73220BFFE7FE97FB10FD88747B" ID-CoL="MTQW" authority="Muller, 1786" authorityName="Muller" authorityYear="1786" box="[373,626,1265,1291]" class="Eurotatoria" family="Brachionidae" genus="Brachionus" kingdom="Animalia" order="Ploima" pageId="7" pageNumber="1991" phylum="Rotifera" rank="species" species="plicatilis">Brachionus plicatilis</taxonomicName>
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</paragraph>
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</subSubSection>
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</emphasis>
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</heading>
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<subSubSection id="C305657B220BFFE9FF42FAC4FE7F7355" lastPageId="9" lastPageNumber="1993" pageId="7" pageNumber="1991" type="description">
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<paragraph id="8BA036F0220BFFE7FF42FAC4FF28744F" blockId="7.[160,1156,1317,1447]" box="[160,210,1317,1343]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FAC4FF28744F" bold="true" box="[160,210,1317,1343]" italics="true" pageId="7" pageNumber="1991">SEM</emphasis>
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FAA8FD5774D7" blockId="7.[160,1156,1317,1447]" pageId="7" pageNumber="1991">
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Diapausing embryos were ovoid and measured ca 134 µm in diameter
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<emphasis id="B96BEAE2220BFFE7FC12FAA8FBFF742F" box="[1008,1029,1350,1378]" italics="true" pageId="7" pageNumber="1991">(̄x</emphasis>
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= 134.0 ± 4.0 μm). The outer surface was composed of numerous smooth ridges and wrinkles, creating an overall rough surface (
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<figureCitation id="13242A75220BFFE7FDC1FA6FFD5A74D7" box="[547,672,1422,1447]" captionStart="Figure 2" captionStartId="8.[160,227,1351,1374]" captionTargetBox="[163,1153,142,1319]" captionTargetId="figure-3@8.[163,1153,142,1319]" captionTargetPageId="8" captionText="Figure 2. Characterisation of diapausing embryos of Brachionus plicatilis. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm." figureDoi="http://doi.org/10.5281/zenodo.10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="7" pageNumber="1991">Figure 2a,b</figureCitation>
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).
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</paragraph>
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<paragraph id="8BA036F0220BFFE7FF42FA37FF287480" blockId="7.[160,1156,1494,1728]" box="[160,210,1494,1520]" pageId="7" pageNumber="1991">
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<emphasis id="B96BEAE2220BFFE7FF42FA37FF287480" bold="true" box="[160,210,1494,1520]" italics="true" pageId="7" pageNumber="1991">TEM</emphasis>
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</paragraph>
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<paragraph id="8BA036F0220BFFE9FF42FA1BFE4F719D" blockId="7.[160,1156,1494,1728]" lastBlockId="9.[160,1156,142,237]" lastPageId="9" lastPageNumber="1993" pageId="7" pageNumber="1991">
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The shell consisted of a thick apical layer and a thin basal layer. Average shell thickness was 2.30 ± 0.22 µm (SD). The outer layer, termed the alveolar layer by
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<bibRefCitation id="EF8E4B01220BFFE7FC10F9FCFECA7728" author="Munuswamy N & Hagiwara A & Murugan G & Hirayama K & Dumont HJ" pageId="7" pageNumber="1991" pagination="219 - 223" refId="ref11496" refString="Munuswamy N, Hagiwara A, Murugan G, Hirayama K, Dumont HJ. 1996. Structural differences between the resting eggs of Brachionus plicatilis and Brachionus rotundiformis (Rotifera, Brachionidae): an electron microscopic study. Hydrobiologia. 318 (3): 219 - 223. doi: 10.1007 / BF 00016683." type="journal article" year="1996">
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||
Munuswamy
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||
<emphasis id="B96BEAE2220BFFE7FF42F9DEFF2F7728" box="[160,213,1599,1624]" italics="true" pageId="7" pageNumber="1991">et al</emphasis>
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. (1996)
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</bibRefCitation>
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, had a wavy texture (
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||
<figureCitation id="13242A75220BFFE7FDDFF9DEFD3B7728" box="[573,705,1599,1624]" captionStart="Figure 2" captionStartId="8.[160,227,1351,1374]" captionTargetBox="[163,1153,142,1319]" captionTargetId="figure-3@8.[163,1153,142,1319]" captionTargetPageId="8" captionText="Figure 2. Characterisation of diapausing embryos of Brachionus plicatilis. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm." figureDoi="http://doi.org/10.5281/zenodo.10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="7" pageNumber="1991">Figure 2c,d</figureCitation>
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). Electron density was highest at the topmost and bottom portions of the alveolar layer, otherwise it had a similar electron density throughout its thickness and contained many ̍empty̾ pockets that were interconnected through electron-dense bridges. The basal layer [s1 of
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<bibRefCitation id="EF8E4B01220BFFE9FC10F946FEDD71D7" author="Munuswamy N & Hagiwara A & Murugan G & Hirayama K & Dumont HJ" lastPageId="9" lastPageNumber="1993" pageId="7" pageNumber="1991" pagination="219 - 223" refId="ref11496" refString="Munuswamy N, Hagiwara A, Murugan G, Hirayama K, Dumont HJ. 1996. Structural differences between the resting eggs of Brachionus plicatilis and Brachionus rotundiformis (Rotifera, Brachionidae): an electron microscopic study. Hydrobiologia. 318 (3): 219 - 223. doi: 10.1007 / BF 00016683." type="journal article" year="1996">
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Munuswamy
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<emphasis id="B96BEAE22205FFE9FF42FF6FFF2B71D7" box="[160,209,142,167]" italics="true" pageId="9" pageNumber="1993">et al</emphasis>
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. (1996)
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</bibRefCitation>
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] had an amorphous appearance that consisted of an electron lucent zone and a more electron-opaque zone below it (
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<figureCitation id="13242A752205FFE9FD28FF50FCC771BA" box="[714,829,177,202]" captionStart="Figure 2" captionStartId="8.[160,227,1351,1374]" captionTargetBox="[163,1153,142,1319]" captionTargetId="figure-3@8.[163,1153,142,1319]" captionTargetPageId="8" captionText="Figure 2. Characterisation of diapausing embryos of Brachionus plicatilis. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm." figureDoi="http://doi.org/10.5281/zenodo.10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="9" pageNumber="1993">Figure 2d</figureCitation>
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). Thin fibres were present throughout both zones.
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</paragraph>
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<caption id="DF6066782204FFE8FF42FAA6FD8E7738" ID-DOI="http://doi.org/10.5281/zenodo.10479033" ID-Zenodo-Dep="10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="8" pageNumber="1992" startId="8.[160,227,1351,1374]" targetBox="[163,1153,142,1319]" targetPageId="8" targetType="figure">
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<paragraph id="8BA036F02204FFE8FF42FAA6FD8E7738" blockId="8.[160,1157,1351,1608]" pageId="8" pageNumber="1992">
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<emphasis id="B96BEAE22204FFE8FF42FAA6FEFB742E" bold="true" box="[160,257,1351,1374]" pageId="8" pageNumber="1992">Figure 2.</emphasis>
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Characterisation of diapausing embryos of
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<taxonomicName id="4C1F4D732204FFE8FD37FAA6FC67742E" authorityName="Muller" authorityYear="1786" box="[725,925,1351,1374]" class="Eurotatoria" family="Brachionidae" genus="Brachionus" kingdom="Animalia" order="Ploima" pageId="8" pageNumber="1992" phylum="Rotifera" rank="species" species="plicatilis">
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<emphasis id="B96BEAE22204FFE8FD37FAA6FC67742E" box="[725,925,1351,1374]" italics="true" pageId="8" pageNumber="1992">Brachionus plicatilis</emphasis>
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</taxonomicName>
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. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm.
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</paragraph>
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</caption>
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<paragraph id="8BA036F02205FFE9FF42FEF9FF2F7042" blockId="9.[160,1156,280,549]" box="[160,213,280,306]" pageId="9" pageNumber="1993">
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||
<emphasis id="B96BEAE22205FFE9FF42FEF9FF2F7042" bold="true" box="[160,213,280,306]" italics="true" pageId="9" pageNumber="1993">AFM</emphasis>
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</paragraph>
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<paragraph id="8BA036F02205FFE9FF42FEDDFE7F7355" blockId="9.[160,1156,280,549]" pageId="9" pageNumber="1993">
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Topography of the outer shell showed rugosity and similar attributes to those in SEM, but with fewer details (
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<figureCitation id="13242A752205FFE9FDF4FEBFFD767007" box="[534,652,350,375]" captionStart="Figure 2" captionStartId="8.[160,227,1351,1374]" captionTargetBox="[163,1153,142,1319]" captionTargetId="figure-3@8.[163,1153,142,1319]" captionTargetPageId="8" captionText="Figure 2. Characterisation of diapausing embryos of Brachionus plicatilis. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm." figureDoi="http://doi.org/10.5281/zenodo.10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="9" pageNumber="1993">Figure 2e</figureCitation>
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). An example force–distance curve that displays how Young̾s modulus values were collected for a single spot on
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<specimenCount id="9D19FD792205FFE9FBBAFE60FF3670CD" count="1" pageId="9" pageNumber="1993" type="egg">one egg</specimenCount>
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is shown in
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<figureCitation id="13242A752205FFE9FE8DFE45FE2370CD" box="[367,473,420,445]" captionStart="Figure 2" captionStartId="8.[160,227,1351,1374]" captionTargetBox="[163,1153,142,1319]" captionTargetId="figure-3@8.[163,1153,142,1319]" captionTargetPageId="8" captionText="Figure 2. Characterisation of diapausing embryos of Brachionus plicatilis. a. Scanning Electron Microscope image of a single egg. b. Close-up of the surface of the egg. c. Ultrastructure of the shell viewed with Transmission Electron Microscopy. d. Close-up of shell ultrastructure. e. Topographic image of the external surface of an egg generated with Atomic Force Microscopy. f. Force–distance (FD) curve generated from a single location on an egg. Parameters for the position of the location on the eggshell are indicated by the cursor positions (software screenshot). Maximum force (MF, green circle) and 20% maximum force (20% MF, orange circle) are indicated on the FD curve. Abbreviations: al, alveolar layer; es, extraembryonic space (section did not include embryo); s1, sublayer. Scale bars: A = 25 µm; B = 2 µm; C = 6.5 µm; D = 1.5 µm." figureDoi="http://doi.org/10.5281/zenodo.10479033" httpUri="https://zenodo.org/record/10479033/files/figure.png" pageId="9" pageNumber="1993">
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||
Figure
|
||
<specimenCount id="9D19FD792205FFE9FE26FE45FE2370CD" box="[452,473,420,445]" count="2" pageId="9" pageNumber="1993" type="generic">2f</specimenCount>
|
||
</figureCitation>
|
||
. The Young̾s modulus ranged from 12.24 to 19.85 MPa with an average of 15.81 ± 2.49 MPa (SD) (
|
||
<tableCitation id="C69D034B2205FFE9FD50FE27FCF170AF" box="[690,779,454,479]" captionStart="Table 1" captionStartId="6.[317,373,142,165]" captionTargetPageId="6" captionText="Table 1. Young̍s modulus (MPa) of elasticity values for diapausing" httpUri="http://table.plazi.org/id/DF606678220AFFE6FEDFFF6FFDD5706A" pageId="9" pageNumber="1993" tableUuid="DF606678220AFFE6FEDFFF6FFDD5706A">Table 1</tableCitation>
|
||
). Hardness values ranged from 1.14 × 10−2 to 5.31 × 10−2 GPa with an average hardness of 1.853 × 10−2 ±1.23 × 10−2 GPa (
|
||
<tableCitation id="C69D034B2205FFE9FEC2FDEDFE837355" box="[288,377,524,549]" captionStart="Table 2" captionStartId="6.[320,376,802,825]" captionTargetBox="[330,985,955,1362]" captionTargetPageId="6" captionText="Table 2. Hardness values (GPa × 10−2) for diapausing embryos of" httpUri="http://table.plazi.org/id/DF606678220AFFE6FEA2FCC3FE2472DE" pageId="9" pageNumber="1993" tableUuid="DF606678220AFFE6FEA2FCC3FE2472DE">Table 2</tableCitation>
|
||
).
|
||
</paragraph>
|
||
</subSubSection>
|
||
</treatment>
|
||
</document> |