Skip to main content
SpringerLink
Log in

Morphological analysis of lamellar structures in mouse type II pneumocytes by quick-freezing and freeze-drying with osmium tetroxide vapor-fixation

  • ORIGINAL PAPER
  • Published:
Medical Molecular Morphology Aims and scope Submit manuscript

Abstract

The lamellar body is a membranous structure periodically laminating in vesicles that is known as the most distinctive feature of type II pneumocytes by conventional preparation methods for transmission electron microscopy. The quick-freezing and freeze-drying method, followed by osmium tetroxide vapor-fixation (QF-FD-OsV), was performed to examine the in situ morphology of the lamellar body in type II pneumocytes of living mouse lungs. Typical lamellar structures were rarely seen in vesicles of the type II pneumocytes, but amorphous components and dispersed stripes were often detected in the vesicles, as revealed by the QF-FD-OsV method. To clarify how the lamellar body was formed during the conventional preparation steps, lung tissues of mice were treated with different fixation procedures, such as immersion-fixation with osmium tetroxide or perfusion-fixation with glutaraldehyde followed by osmium tetroxide, in combination with alcohol dehydration or QF-FD-OsV. In addition to lamellar bodies of type II pneumocytes in the specimens with alcohol dehydration, some lamellar structures were also formed even with the QF-FD-OsV method. These findings suggest that the labile lamellar body is easily modified and formed during both chemical fixation and alcohol dehydration steps.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. L Weiss RO Greep (1977) Alveolar cells Histology EditionNumber4th edn McGraw-Hill New York 805–811

    Google Scholar 

  2. AW Ham DH Cormack (1979) The pulmonary surfactant Histology EditionNumber8th edn Lippincott Philadelphia 744–748

    Google Scholar 

  3. DW Fawcett (1986) Respiratory system Bloom and Fawcett: Textbook of histology EditionNumber11th edn Saunders Philadelphia 731–754

    Google Scholar 

  4. FB Askin C Kuhn (1971) ArticleTitleThe cellular origin of pulmonary surfactant Lab Invest 25 260–268 Occurrence Handle5109934 Occurrence Handle1:CAS:528:DyaE3MXlt1Olsrc%3D

    PubMed  CAS  Google Scholar 

  5. A Chander AB Fisher (1990) ArticleTitleRegulation of lung surfactant secretion Am J Physiol Lung Cell Mol Physiol 258 241–253

    Google Scholar 

  6. SL Young EK Fram CL Spain EW Larson (1991) ArticleTitleDevelopment of type II pneumocytes in rat lung Am J Physiol 260 113–122

    Google Scholar 

  7. G Schmitz G Muller (1991) ArticleTitleStructure and function of lamellar bodies, lipid-protein complexes involved in storage and secretion of cellular lipids J Lipid Res 32 1539–1570 Occurrence Handle1797938 Occurrence Handle1:CAS:528:DyaK3MXmslKgsbc%3D

    PubMed  CAS  Google Scholar 

  8. CJ Stratton (1975) ArticleTitleMultilamellar body formation in mammalian lung: an ultrastructural study utilizing three lipid-retention procedures J Ultrastruct Res 52 309–320 Occurrence Handle51093 Occurrence Handle1:STN:280:DyaE28%2FgslOgug%3D%3D Occurrence Handle10.1016/S0022-5320(75)80071-2

    Article  PubMed  CAS  Google Scholar 

  9. CJ Stratton (1976) ArticleTitleThe high resolution ultrastructural of the periodicity and architecture of lipid-retained and extracted lung multilamellar body laminations Tissue Cell 8 713–728 Occurrence Handle828331 Occurrence Handle1:STN:280:DyaE2s7ivVGnsg%3D%3D Occurrence Handle10.1016/0040-8166(76)90041-0

    Article  PubMed  CAS  Google Scholar 

  10. RJ Sanderson AE Vatter (1977) ArticleTitleA mode of formation of tubular myelin from lamellar bodies in the lung J Cell Biol 74 1027–1031 Occurrence Handle903368 Occurrence Handle1:STN:280:DyaE1c%2FgsFKisQ%3D%3D Occurrence Handle10.1083/jcb.74.3.1027

    Article  PubMed  CAS  Google Scholar 

  11. MC Williams (1978) ArticleTitleFreeze-fracture studies of tubular myelin and lamellar bodies in fetal and adult rat lungs J Ultrastruct Res 64 352–361 Occurrence Handle581389 Occurrence Handle1:STN:280:DyaE1M%2FltVKksQ%3D%3D Occurrence Handle10.1016/S0022-5320(78)90043-6

    Article  PubMed  CAS  Google Scholar 

  12. AJ Collet (1979) ArticleTitlePreservation of alveolar type II pneumocyte lamellar bodies for electron microscopic studies J Histochem Cytochem 27 989–996 Occurrence Handle113451 Occurrence Handle1:STN:280:DyaL3c%2FgslCrsQ%3D%3D

    PubMed  CAS  Google Scholar 

  13. CJ Stratton TB Erickson HY Wetzstein (1982) ArticleTitleThe lipid solubility of fixative, staining and embedding media, and the introduction of LX-112 and poly/bed-812 as dehydrants for expoxy resin embedment Tissue Cell 14 13–24 Occurrence Handle7046136 Occurrence Handle1:CAS:528:DyaL38Xlt1eitLk%3D Occurrence Handle10.1016/0040-8166(82)90003-9

    Article  PubMed  CAS  Google Scholar 

  14. WHJ Douglas SA Redding M Stain (1975) ArticleTitleThe lamellar substructure of osmiophilic inclusion bodies present in rat type II alveolar pneumocytes Tissue Cell 7 137–142 Occurrence Handle164061 Occurrence Handle1:STN:280:DyaE2M7it1Ohtw%3D%3D Occurrence Handle10.1016/S0040-8166(75)80011-5

    Article  PubMed  CAS  Google Scholar 

  15. K Saga (2005) ArticleTitleApplication of cryofixation and cryoultramicrotomy for biological electron microscopy Med Mol Morphol 38 155–160 Occurrence Handle16170463 Occurrence Handle10.1007/s00795-005-0287-2

    Article  PubMed  Google Scholar 

  16. HD Coulter L Terracio (1977) ArticleTitlePreparation of biological tissue for electron microscopy by freeze-drying Anat Rec 187 477–494 Occurrence Handle848780 Occurrence Handle1:STN:280:DyaE2s7lsFeksw%3D%3D Occurrence Handle10.1002/ar.1091870405

    Article  PubMed  CAS  Google Scholar 

  17. SL Erlandsen JA Parson JP Burke DE Van Orden LS Van Orden (1975) ArticleTitleA modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagons, and growth hormone J Histochem Cytochem 23 666–677 Occurrence Handle1176760 Occurrence Handle1:STN:280:DyaE28%2FisFylsQ%3D%3D

    PubMed  CAS  Google Scholar 

  18. RW Dudek GV Childs AF Boyne (1982) ArticleTitleQuick-freezing and freeze-drying in preparation for high quality morphology and immunocytochemistry at the ultrastructural level: application to pancreatic beta cell J Histochem Cytochem 30 129–138 Occurrence Handle7037935 Occurrence Handle1:STN:280:DyaL387ktlGltg%3D%3D

    PubMed  CAS  Google Scholar 

  19. HD Coulter RP Elde (1978) ArticleTitleSomatostatin radioimmunoassay and immunofluorescence in the rat hypothalamus: effects of dehydration with alcohol and fixation with aldehydes and OsO4 Anat Rec 190 369–370

    Google Scholar 

  20. L Terracio JA McAteer (1982) ArticleTitleLamellar body structure in cultured fetal rat type II cells as demonstrated by rapid-freezing and freeze-drying Anat Rec 204 404–405

    Google Scholar 

  21. L Terracio JA McAteer (1984) ArticleTitlePulmonary type II cell lamellar body ultrastructure preserved by rapid freezing and freeze drying Anat Rec 209 355–362 Occurrence Handle6465544 Occurrence Handle10.1002/ar.1092090314

    Article  PubMed  Google Scholar 

  22. S Ohno N Terada Y Fujii H Ueda I Takayama (1996) ArticleTitleDynamic structure of glomerular capillary loop as revealed by an in vivo cryotechnique Virchows Arch 427 519–527 Occurrence Handle8624582 Occurrence Handle1:STN:280:DyaK283htFOjsA%3D%3D Occurrence Handle10.1007/BF00199513

    Article  PubMed  CAS  Google Scholar 

  23. N Ohno N Terada S Ohno (2004) ArticleTitleAdvanced application of the in vivo cryotechnique to immunohistochemistry for animal organs Acta Histochem Cytochem 37 357–364 Occurrence Handle10.1267/ahc.37.357

    Article  Google Scholar 

  24. Z Li N Terada N Ohno S Ohno (2005) ArticleTitleImmunohistochemical analyses on albumin and immunoglobulin in acute hypertensive mouse kidneys by “in vivo cryotechnique Histol Histopathol 20 807–816 Occurrence Handle15944930 Occurrence Handle1:CAS:528:DC%2BD2MXpsFKjt74%3D

    PubMed  CAS  Google Scholar 

  25. N Terada N Ohno Z Li Y Fujii T Baba S Ohno (2005) ArticleTitleDetection of injected fluorescence-conjugated IgG in living mouse organs using “in vivo cryotechnique” with freeze-substitution Microsc Res Tech 66 173–178 Occurrence Handle15889425 Occurrence Handle1:CAS:528:DC%2BD2MXlvVCrurc%3D Occurrence Handle10.1002/jemt.20155

    Article  PubMed  CAS  Google Scholar 

  26. Ohno N, Terada N, Ohno S (2006) Histochemical analyses of living mouse liver under different hemodynamic conditions by “in vivo cryotechnique.” Histochem Cell Biol (in press)

  27. I Takayama N Terada T Baba H Ueda Y Fujii Y Kato S Ohno (2000) ArticleTitleDynamic ultrastructure of mouse pulmonary alveoli revealed by an in vivo cryotechnique in combination with freeze-substitution J Anat 197 199–205 Occurrence Handle11005712 Occurrence Handle10.1046/j.1469-7580.2000.19720199.x

    Article  PubMed  Google Scholar 

  28. N Ohno N Terada S Murata R Katoh S Ohno (2005) ArticleTitleApplication of cryotechniques with freeze-substitution for the immunohistochemical demonstration of intranuclear pCREB and chromosome territory J Histochem Cytochem 53 55–62 Occurrence Handle15637338 Occurrence Handle1:CAS:528:DC%2BD2MXot12gsQ%3D%3D Occurrence Handle10.1369/jhc.4A6343.2005

    Article  PubMed  CAS  Google Scholar 

  29. N Terada N Ohno Z Li Y Fujii T Baba S Ohno (2006) ArticleTitleApplication of in vivo cryotechnique to the examination of cells and tissues in living animal organs Histol Histopathol 21 265–272 Occurrence Handle16372248 Occurrence Handle1:STN:280:DC%2BD2MnotlaksA%3D%3D

    PubMed  CAS  Google Scholar 

  30. PM Frederik PH Bomans WM Busing R Odselius WM Hax (1982) ArticleTitleVapor fixation for immunocytochemistry and X-ray microanalysis on cryoultramicrotome sections J Histochem Cytochem 32 636–642

    Google Scholar 

  31. WJ Martin DL Kachel (1987) ArticleTitleIprindole reverses the lamellar body deficiency of cultured L-2 cells, possible implications in the reverses of surfactant deficiency Am J Pathol 129 34–43 Occurrence Handle2821815 Occurrence Handle1:CAS:528:DyaL1cXjtFKrsw%3D%3D

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 1110 Shimokato, Chuo-shi, Yamanashi, 409-3898, Japan

    Chunlu Yang, Nobuo Terada, Nobuhiko Ohno, Yasuhisa Fujii & Shinichi Ohno

  2. Department of Thoracic Surgery, First Hospital of China Medical University, Shenyang, P. R. China

    Chunlu Yang

Authors
  1. Chunlu Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nobuo Terada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nobuhiko Ohno
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yasuhisa Fujii
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shinichi Ohno
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shinichi Ohno.

Additional information

Dr. C. Yang was a foreign research fellow from the Department of Thoracic Surgery, the First Hospital of China Medical University, Shenyang, P. R. China, while this work was in progress at the University of Yamanashi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, C., Terada, N., Ohno, N. et al. Morphological analysis of lamellar structures in mouse type II pneumocytes by quick-freezing and freeze-drying with osmium tetroxide vapor-fixation. Med Mol Morphol 39, 88–96 (2006). https://doi.org/10.1007/s00795-006-0318-7

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00795-006-0318-7

Key words

Search

Navigation