DOI: 10.1111/j.1365-2818.2006.01558.x. Aclar discs: a versatile substrate for routine high‐pressure freezing of mammalian cell monolayers

Aclar discs: a versatile substrate for routine high‐pressure freezing of mammalian cell monolayers

10.1111/j.1365-2818.2006.01558.x

Crossref journal-article

Wiley

Journal of Microscopy (311)

Abstract

SummaryHigh‐pressure freezing avoids the artefacts induced by conventional chemical fixation, and, in combination with freeze‐substitution and plastic embedding, is a reliable method for the ultrastructural analysis of mammalian cell monolayers. In order to high‐pressure freeze mammalian cell monolayers, cells have to be seeded on a suitable substrate. Unfortunately, electron microscopy analysis is often hampered by poor cell growth, changes in cell morphology induced by the cell substrate or cell loss during processing. We report a method to culture, high‐pressure freeze, freeze‐substitute and plastic embed mammalian cell monolayers. The method is based on the use of Aclar, a copolymer film with properties very similar to those of tissue culture plastic. We show that Aclar discs support the normal growth and morphology of a wide variety of mammalian cell types, and form an ideal starting point for high‐pressure freezing, freeze‐substitution and plastic embedding. We present a complete protocol, which, because of its simplicity and reproducibility, provides a method suitable for the routine analysis of mammalian cell monolayers by electron microscopy and tomography.

Bibliography

JIMÃNEZ, N., HUMBEL, B. M., VAN DONSELAAR, E., VERKLEIJ, A. J., & BURGER, K. N. J. (2006). Aclar discs: a versatile substrate for routine highâpressure freezing of mammalian cell monolayers. Journal of Microscopy, 221(3), 216â223. Portico.

Authors 5

N. JIMÉNEZ (first)
B. M. HUMBEL (additional)
E. VAN DONSELAAR (additional)
A. J. VERKLEIJ (additional)
K. N. J. BURGER (additional)

References 28 Referenced 41

10.1046/j.1365-2818.2002.01051.x
10.1007/BF00495382
10.3732/ajb.91.3.313
10.1006/jmcc.1997.0408
{'key': 'e_1_2_5_6_1', 'first-page': '715', 'volume-title': 'Protein–Protein Interactions: a Molecular Cloning Manual', 'author': 'Geerts W.J.C.', 'year': '2005'} / Protein–Protein Interactions: a Molecular Cloning Manual by Geerts W.J.C. (2005)
10.1002/jemt.1060030206
10.1006/jsbi.2000.4230
{'key': 'e_1_2_5_9_1', 'first-page': '175', 'volume-title': 'The Science of Biological Specimen Preparation, 1985', 'author': 'Humbel B.M.', 'year': '1986'} / The Science of Biological Specimen Preparation, 1985 by Humbel B.M. (1986)
10.1002/jemt.1060100110
10.1093/emboj/20.20.5636
10.1083/jcb.144.6.1135
10.1073/pnas.051631998
10.1006/jsbi.1997.3919
10.3109/10520296809115061
10.1097/00005072-197107000-00001
10.1002/jemt.1060120211
10.1007/s004120050320
10.1023/A:1008894612021
10.1046/j.1365-2818.2003.01231.x
{'key': 'e_1_2_5_21_1', 'first-page': '131', 'volume-title': 'Science of Biological Specimen Preparation, 1983', 'author': 'Muller M.', 'year': '1984'} / Science of Biological Specimen Preparation, 1983 by Muller M. (1984)
10.1046/j.1365-2818.2002.01085.x
10.1046/j.1365-2818.2003.01225.x
{'key': 'e_1_2_5_24_1', 'volume-title': 'Morphologische grundlagen zur zell‐zell interaktion bei adulten herzmuskelzellen in kultur.', 'author': 'Schwarb P.', 'year': '1990'} / Morphologische grundlagen zur zell‐zell interaktion bei adulten herzmuskelzellen in kultur. by Schwarb P. (1990)
10.1046/j.1365-2818.2001.00919.x
10.1083/jcb.25.1.117
10.1046/j.1365-2818.2002.01064.x
10.1002/jemt.1098
10.1046/j.0022-2720.2001.00987.x

Dates

Type	When
Created	19 years, 5 months ago (March 21, 2006, 9:51 a.m.)
Deposited	1 year, 10 months ago (Oct. 12, 2023, 3:30 p.m.)
Indexed	4 months, 2 weeks ago (April 7, 2025, 5:03 a.m.)
Issued	19 years, 5 months ago (March 1, 2006)
Published	19 years, 5 months ago (March 1, 2006)
Published Online	19 years, 5 months ago (March 21, 2006)
Published Print	19 years, 5 months ago (March 1, 2006)

Funders 0

None

BibTeX

@article{JIM_NEZ_2006, title={Aclar discs: a versatile substrate for routine high‐pressure freezing of mammalian cell monolayers}, volume={221}, ISSN={1365-2818}, url={http://dx.doi.org/10.1111/j.1365-2818.2006.01558.x}, DOI={10.1111/j.1365-2818.2006.01558.x}, number={3}, journal={Journal of Microscopy}, publisher={Wiley}, author={JIMÉNEZ, N. and HUMBEL, B. M. and VAN DONSELAAR, E. and VERKLEIJ, A. J. and BURGER, K. N. J.}, year={2006}, month=mar, pages={216–223} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        4,
        7
      ]
    ],
    "date-time": "2025-04-07T09:03:07Z",
    "timestamp": 1744016587635
  },
  "reference-count": 28,
  "publisher": "Wiley",
  "issue": "3",
  "license": [
    {
      "start": {
        "date-parts": [
          [
            2006,
            3,
            21
          ]
        ],
        "date-time": "2006-03-21T00:00:00Z",
        "timestamp": 1142899200000
      },
      "content-version": "vor",
      "delay-in-days": 20,
      "URL": "http://onlinelibrary.wiley.com/termsAndConditions#vor"
    }
  ],
  "content-domain": {
    "domain": [
      "onlinelibrary.wiley.com"
    ],
    "crossmark-restriction": true
  },
  "published-print": {
    "date-parts": [
      [
        2006,
        3
      ]
    ]
  },
  "abstract": "<jats:title>Summary</jats:title><jats:p>High\u2010pressure freezing avoids the artefacts induced by conventional chemical fixation, and, in combination with freeze\u2010substitution and plastic embedding, is a reliable method for the ultrastructural analysis of mammalian cell monolayers. In order to high\u2010pressure freeze mammalian cell monolayers, cells have to be seeded on a suitable substrate. Unfortunately, electron microscopy analysis is often hampered by poor cell growth, changes in cell morphology induced by the cell substrate or cell loss during processing. We report a method to culture, high\u2010pressure freeze, freeze\u2010substitute and plastic embed mammalian cell monolayers. The method is based on the use of Aclar, a copolymer film with properties very similar to those of tissue culture plastic. We show that Aclar discs support the normal growth and morphology of a wide variety of mammalian cell types, and form an ideal starting point for high\u2010pressure freezing, freeze\u2010substitution and plastic embedding. We present a complete protocol, which, because of its simplicity and reproducibility, provides a method suitable for the routine analysis of mammalian cell monolayers by electron microscopy and tomography.</jats:p>",
  "DOI": "10.1111/j.1365-2818.2006.01558.x",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2006,
        3,
        21
      ]
    ],
    "date-time": "2006-03-21T14:51:27Z",
    "timestamp": 1142952687000
  },
  "page": "216-223",
  "update-policy": "http://dx.doi.org/10.1002/crossmark_policy",
  "source": "Crossref",
  "is-referenced-by-count": 41,
  "title": "Aclar discs: a versatile substrate for routine high\u2010pressure freezing of mammalian cell monolayers",
  "prefix": "10.1111",
  "volume": "221",
  "author": [
    {
      "given": "N.",
      "family": "JIM\u00c9NEZ",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "B. M.",
      "family": "HUMBEL",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "E.",
      "family": "VAN DONSELAAR",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "A. J.",
      "family": "VERKLEIJ",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "K. N. J.",
      "family": "BURGER",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "311",
  "published-online": {
    "date-parts": [
      [
        2006,
        3,
        21
      ]
    ]
  },
  "reference": [
    {
      "key": "e_1_2_5_2_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2002.01051.x"
    },
    {
      "key": "e_1_2_5_3_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/BF00495382"
    },
    {
      "key": "e_1_2_5_4_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.3732/ajb.91.3.313"
    },
    {
      "key": "e_1_2_5_5_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jmcc.1997.0408"
    },
    {
      "key": "e_1_2_5_6_1",
      "first-page": "715",
      "volume-title": "Protein\u2013Protein Interactions: a Molecular Cloning Manual",
      "author": "Geerts W.J.C.",
      "year": "2005"
    },
    {
      "key": "e_1_2_5_7_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/jemt.1060030206"
    },
    {
      "key": "e_1_2_5_8_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jsbi.2000.4230"
    },
    {
      "key": "e_1_2_5_9_1",
      "first-page": "175",
      "volume-title": "The Science of Biological Specimen Preparation, 1985",
      "author": "Humbel B.M.",
      "year": "1986"
    },
    {
      "key": "e_1_2_5_10_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/jemt.1060100110"
    },
    {
      "key": "e_1_2_5_11_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/emboj/20.20.5636"
    },
    {
      "key": "e_1_2_5_12_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.144.6.1135"
    },
    {
      "key": "e_1_2_5_13_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.051631998"
    },
    {
      "key": "e_1_2_5_14_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1006/jsbi.1997.3919"
    },
    {
      "key": "e_1_2_5_15_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.3109/10520296809115061"
    },
    {
      "key": "e_1_2_5_16_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1097/00005072-197107000-00001"
    },
    {
      "key": "e_1_2_5_17_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/jemt.1060120211"
    },
    {
      "key": "e_1_2_5_18_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1007/s004120050320"
    },
    {
      "key": "e_1_2_5_19_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1023/A:1008894612021"
    },
    {
      "key": "e_1_2_5_20_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2003.01231.x"
    },
    {
      "key": "e_1_2_5_21_1",
      "first-page": "131",
      "volume-title": "Science of Biological Specimen Preparation, 1983",
      "author": "Muller M.",
      "year": "1984"
    },
    {
      "key": "e_1_2_5_22_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2002.01085.x"
    },
    {
      "key": "e_1_2_5_23_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2003.01225.x"
    },
    {
      "key": "e_1_2_5_24_1",
      "volume-title": "Morphologische grundlagen zur zell\u2010zell interaktion bei adulten herzmuskelzellen in kultur.",
      "author": "Schwarb P.",
      "year": "1990"
    },
    {
      "key": "e_1_2_5_25_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2001.00919.x"
    },
    {
      "key": "e_1_2_5_26_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.25.1.117"
    },
    {
      "key": "e_1_2_5_27_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.1365-2818.2002.01064.x"
    },
    {
      "key": "e_1_2_5_28_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1002/jemt.1098"
    },
    {
      "key": "e_1_2_5_29_1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1046/j.0022-2720.2001.00987.x"
    }
  ],
  "container-title": "Journal of Microscopy",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1365-2818.2006.01558.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "text-mining"
    },
    {
      "URL": "https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2818.2006.01558.x",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2023,
        10,
        12
      ]
    ],
    "date-time": "2023-10-12T19:30:26Z",
    "timestamp": 1697139026000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://onlinelibrary.wiley.com/doi/10.1111/j.1365-2818.2006.01558.x"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        2006,
        3
      ]
    ]
  },
  "references-count": 28,
  "journal-issue": {
    "issue": "3",
    "published-print": {
      "date-parts": [
        [
          2006,
          3
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1111/j.1365-2818.2006.01558.x"
  ],
  "URL": "http://dx.doi.org/10.1111/j.1365-2818.2006.01558.x",
  "relation": {},
  "ISSN": [
    "0022-2720",
    "1365-2818"
  ],
  "subject": [],
  "container-title-short": "Journal of Microscopy",
  "published": {
    "date-parts": [
      [
        2006,
        3
      ]
    ]
  },
  "assertion": [
    {
      "value": "2006-03-21",
      "order": 2,
      "name": "published",
      "label": "Published",
      "group": {
        "name": "publication_history",
        "label": "Publication History"
      }
    }
  ]
}