Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy

Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protoco...
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Autor*in:	Kim, Seong-Oh [verfasserIn] Kim, Joonhui Okajima, Takaharu Cho, Nam-Joon

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2017

Schlagwörter:	Scanning ion conductance microscopy Atomic force microscopy Cellular mechanics Cell fixation

Anmerkung:	© Korea Nano Technology Research Society 2017

Übergeordnetes Werk:	Enthalten in: Nano Convergence - Berlin : SpringerOpen, 2014, 4(2017), 1 vom: 20. März
Übergeordnetes Werk:	volume:4 ; year:2017 ; number:1 ; day:20 ; month:03

Links:	Volltext

DOI / URN:	10.1186/s40580-017-0099-9

Katalog-ID:	SPR036686883

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520			\|a Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols.
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allfields	10.1186/s40580-017-0099-9 doi (DE-627)SPR036686883 (SPR)s40580-017-0099-9-e DE-627 ger DE-627 rakwb eng Kim, Seong-Oh verfasserin aut Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Korea Nano Technology Research Society 2017 Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213 Kim, Joonhui aut Okajima, Takaharu aut Cho, Nam-Joon aut Enthalten in Nano Convergence Berlin : SpringerOpen, 2014 4(2017), 1 vom: 20. März (DE-627)780378938 (DE-600)2760386-6 2196-5404 nnns volume:4 year:2017 number:1 day:20 month:03 https://dx.doi.org/10.1186/s40580-017-0099-9 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2017 1 20 03
spelling	10.1186/s40580-017-0099-9 doi (DE-627)SPR036686883 (SPR)s40580-017-0099-9-e DE-627 ger DE-627 rakwb eng Kim, Seong-Oh verfasserin aut Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Korea Nano Technology Research Society 2017 Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213 Kim, Joonhui aut Okajima, Takaharu aut Cho, Nam-Joon aut Enthalten in Nano Convergence Berlin : SpringerOpen, 2014 4(2017), 1 vom: 20. März (DE-627)780378938 (DE-600)2760386-6 2196-5404 nnns volume:4 year:2017 number:1 day:20 month:03 https://dx.doi.org/10.1186/s40580-017-0099-9 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2017 1 20 03
allfields_unstemmed	10.1186/s40580-017-0099-9 doi (DE-627)SPR036686883 (SPR)s40580-017-0099-9-e DE-627 ger DE-627 rakwb eng Kim, Seong-Oh verfasserin aut Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Korea Nano Technology Research Society 2017 Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213 Kim, Joonhui aut Okajima, Takaharu aut Cho, Nam-Joon aut Enthalten in Nano Convergence Berlin : SpringerOpen, 2014 4(2017), 1 vom: 20. März (DE-627)780378938 (DE-600)2760386-6 2196-5404 nnns volume:4 year:2017 number:1 day:20 month:03 https://dx.doi.org/10.1186/s40580-017-0099-9 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2017 1 20 03
allfieldsGer	10.1186/s40580-017-0099-9 doi (DE-627)SPR036686883 (SPR)s40580-017-0099-9-e DE-627 ger DE-627 rakwb eng Kim, Seong-Oh verfasserin aut Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Korea Nano Technology Research Society 2017 Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213 Kim, Joonhui aut Okajima, Takaharu aut Cho, Nam-Joon aut Enthalten in Nano Convergence Berlin : SpringerOpen, 2014 4(2017), 1 vom: 20. März (DE-627)780378938 (DE-600)2760386-6 2196-5404 nnns volume:4 year:2017 number:1 day:20 month:03 https://dx.doi.org/10.1186/s40580-017-0099-9 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2017 1 20 03
allfieldsSound	10.1186/s40580-017-0099-9 doi (DE-627)SPR036686883 (SPR)s40580-017-0099-9-e DE-627 ger DE-627 rakwb eng Kim, Seong-Oh verfasserin aut Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy 2017 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier © Korea Nano Technology Research Society 2017 Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213 Kim, Joonhui aut Okajima, Takaharu aut Cho, Nam-Joon aut Enthalten in Nano Convergence Berlin : SpringerOpen, 2014 4(2017), 1 vom: 20. März (DE-627)780378938 (DE-600)2760386-6 2196-5404 nnns volume:4 year:2017 number:1 day:20 month:03 https://dx.doi.org/10.1186/s40580-017-0099-9 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4335 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2017 1 20 03
language	English
source	Enthalten in Nano Convergence 4(2017), 1 vom: 20. März volume:4 year:2017 number:1 day:20 month:03
sourceStr	Enthalten in Nano Convergence 4(2017), 1 vom: 20. März volume:4 year:2017 number:1 day:20 month:03
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Scanning ion conductance microscopy Atomic force microscopy Cellular mechanics Cell fixation
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container_title	Nano Convergence
authorswithroles_txt_mv	Kim, Seong-Oh @@aut@@ Kim, Joonhui @@aut@@ Okajima, Takaharu @@aut@@ Cho, Nam-Joon @@aut@@
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Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. 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author	Kim, Seong-Oh
spellingShingle	Kim, Seong-Oh misc Scanning ion conductance microscopy misc Atomic force microscopy misc Cellular mechanics misc Cell fixation Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
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topic_title	Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy Scanning ion conductance microscopy (dpeaa)DE-He213 Atomic force microscopy (dpeaa)DE-He213 Cellular mechanics (dpeaa)DE-He213 Cell fixation (dpeaa)DE-He213
topic	misc Scanning ion conductance microscopy misc Atomic force microscopy misc Cellular mechanics misc Cell fixation
topic_unstemmed	misc Scanning ion conductance microscopy misc Atomic force microscopy misc Cellular mechanics misc Cell fixation
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title	Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
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title_full	Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
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author_browse	Kim, Seong-Oh Kim, Joonhui Okajima, Takaharu Cho, Nam-Joon
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title_sort	mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
title_auth	Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
abstract	Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. © Korea Nano Technology Research Society 2017
abstractGer	Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. © Korea Nano Technology Research Society 2017
abstract_unstemmed	Background Cell fixation is an essential step to preserve cell samples for a wide range of biological assays involving histochemical and cytochemical analysis. Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. Conclusions Taken together, these findings offer a deeper understanding of how varying PFA concentrations influence the mechanical properties of cells and suggest new avenues for establishing refined cell fixation protocols. © Korea Nano Technology Research Society 2017
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title_short	Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy
url	https://dx.doi.org/10.1186/s40580-017-0099-9
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author2	Kim, Joonhui Okajima, Takaharu Cho, Nam-Joon
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Paraformaldehyde (PFA) has been widely used as a cross-linking fixation agent. It has been empirically recognized in a gold standard protocol that the PFA concentration for cell fixation, CPFA, is 4%. However, it is still not quantitatively clear how the conventional protocol of CPFA is optimized. Methods Here, we investigated the mechanical properties of cell fixation as a function of CPFA by using atomic force microscopy and scanning ion conductance microscopy. The goal of this study is to investigate the effect of CPFA (0–10 wt%) on the morphological and mechanical properties of live and fixed mouse fibroblast cells. Results We found that both Young’s modulus, E, and the fluctuation amplitude of apical cell membrane, am, were almost constant in a lower CPFA (<$ 10^{−4} $%). Interestingly, in an intermediate CPFA between $ 10^{−1} $ and 4%, E dramatically increased whereas am abruptly decreased, indicating that entire cells begin to fix at CPFA = ca. $ 10^{−1} $%. Moreover, these quantities were unchanged in a higher CPFA (>4%), indicating that the cell fixation is stabilized at CPFA = ca. 4%, which is consistent with the empirical concentration of cell fixation optimized in biological protocols. 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Mechanical properties of paraformaldehyde-treated individual cells investigated by atomic force microscopy and scanning ion conductance microscopy

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